Cell Signalling Notes Merit Life Sciences1

MERIT LIFE SCIENCES ONLINE Coaching for CSIR NET JRF LIFE SCEINCES
UNIT 4 MERIT LIFE SCIENCES

Cell signalling
As living organisms we are constantly receiving and interpreting signals from our
environment. These signals can be light, heat, odours, touch or sound. The cells of our
bodies are also constantly receiving signals from other cells. These signals are important
to keep cells alive and functioning as well as to stimulate important events such as cell
division and differentiation. In animals, rapid responses to the changes in the
environment are mediated primarily by the nervous system and by hormones including
small peptides, small nonpeptide molecules such as the catecholamines (Dopamine,
epinephrine, norepinephrine). We have already studied role of hormones like
epinephrine, ACTH and norepinephrine etc in signaling events. We shall discuss
downstream process when signaling molecule interacts with receptor. We shall also talk
about various secondary messengers involved in signaling process. These signaling
molecules are released from the cells and they travel through the blood to their specific
target cells as shown in Figure 1. Some molecules are transported long distances by the
blood while others have more of local effects. Certain membrane-bound proteins on one
cell can directly signal an adjacent cell.
Figure 1: Signaling molecules released from cell and transported by the blood to the target cell
Page 1 of 61
NPTEL – Biotechnology – Cell Biology
Cell signaling can be divided into 3 stages:
1. Reception: A cell detects a signaling molecule from the outside of the cell. A signal is
detected when the ligand binds to a receptor protein on the surface of the cell or inside
the cell.
2. Transduction: When the signaling molecule binds to the receptor, it changes the
receptor protein. This change initiates the process of transduction. Each relay molecule in
the signal transduction pathway changes the next molecule in the pathway.
3. Response: Finally, the signal triggers a specific cellular response as shown in Figure 2.
Extracellular fluid Cytoplasm
1. Reception 2. Transduction 3. Response

Cellular
respons
e
Relay molecules
Signaling Plasma
molecule membran
e
Figure 2: Cell signalling stages

Signal Transduction:
Signal transduction is phenomenon which involves in the transfer of signal from
extracellular to intracellular environment through the cell surface receptor protein that
stimulate intracellular target enzymes, which may be either directly linked or indirectly
coupled to receptors by G proteins. These intracellular enzymes serve as downstream
signalling elements that propagate and amplify the signal initiated by ligand binding.
Thus, signal transduction pathway allows cells to respond to extracellular environmental
signals. These signals can be physical and chemical such as light, oxygen, nutrient,
hormones. Figure 3 represents the signal transduction pathway.
Signal transduction is the combination of following phenomenon:
1. Signal reception
2. Integration
3. Amplification
4. A target that is affected
5. Termination
Terminatio
Signals
Figure 3: Representation of Signal Transduction pathway
Thus signal transduction begins with receiving signal to the cell receptor and end with a
change in cellular function. The cell receptor can be of various types- G-protein coupled
receptor, tyrosin kinase receptor etc. The transduction process is typically mediated via a
cascade of some important second messengers including cAMP, cGMP, calcium ion,
inositol 1, 4, 5-trisphosphate, (IP3), and diacylglycerol (DAG). Second messengers are
intracellular molecules that change in concentration in response to environmental signals
and involve in conveying information inside the cell.
Cyclic GMP
Figure 4: Common second messengers
Signal transduction pathways act similar to molecular circuit. This pathway depends on
following factors during transformation of signal from extracellular environment to
intracellular.
1. Signal reception by cell membrane receptor: Some non polar signaling molecules such
as estrogens and other steroid hormones are able to cross the bilipid membrane and hence
make entry inside the cell. Once inside the cell, these molecules can bind to proteins that
interact directly with DNA and involve in regulation of gene transcription. Thus, a
chemical signal enters the cell and directly alters gene-expression patterns. However,
most of signalling molecules are too large and too polar so they are unable to cross the
membrane, hence there is no appropriate transport system. In this case these signaling
molecules transmit signals through cell surface receptor protein without crossing the cell
membrane. We will discuss about cell receptors in upcoming lecture notes.
These receptors are intrinsic membrane protein which consist both extracellular and
intracellular domain. A binding site present in extracellular domain specifically
recognizes the signaling molecule (i.e. well known as ligand). Such binding sites are
analogous to enzyme active sites except that no catalysis takes place within them. When
these signal molecules comes and bind to binding site on receptor protein in extracellular
region then some conformational change occurs in tertiary and quaternary structure of the
receptor which results in the drastic change in the intracellular domain of the receptor.
These structural changes are not sufficient to yield an appropriate response, because they
are restricted to a small number of receptor molecules in the cell membrane. The
information embodied by the presence of the ligand, often called the primary messenger,
must be transduced into other forms that can alter the biochemistry of the cell.
2. Second messengers: Second messengers act as the intermediate molecule that relay
signals from receptors on cell surface to target molecule inside cells, in cytoplasm or
nucleus.
The use of second messengers has several consequences:
a) The second messengers are able to diffuse frequently into other compartment of the
cell such as nucleus where they can influence gene expression and other process.
b) Generation of second messengers leads to amplification of signal. Each signaling
molecule is involved in the generation of several second messengers in the cell. Thus,
a low concentration of signal in the environment, even as little as a single molecule,
can yield a large intracellular signal and response
c) Since common second messengers generate in different signaling pathway, thus the
coordination of signal transduction is driven by interaction between these pathways.
Multiple signaling pathways create both opportunities and potential problems.
Interactions between signaling pathways enables the cell to process and interpret
multiple inputs differently in different contexts leading to cross-talk. Cross talk
between second messengers cause oscillation of various second messengers and also
creates biostability between two steady states. Thus cross talk more precisely involves
in regulation of cell activity than individual independent pathways without cross talk.
However, inappropriate cross-talk can cause second messengers to be misinterpreted.
3. Protein phosphorylation: Protein phosphorylation is most common route for
transferring information coming through second messenger which involve elicit
responses by activating protein kinases. Protein phosphorylation is a post-
translational modification of proteins by phosphorylation at serine, threonine or
tyrosine residues by a protein kinase by the addition of a covalently bound phosphate
group from ATP.
Figure 5: Action of cAMP-dependent protein kinase
4. Signal termination by protein phosphatase: Signal termination is final step of signal

transduction. The well known route for signal termination is by protein phosphatase
enzyme. The signalling process must be terminated after signaling process has been
initiated and the information has been transduced to affect other cellular processes,
because without such termination cells lose their responsiveness to new signals.
Additionally, if termination fails in signaling processes, it may lead to uncontrolled
cell growth and thus increases the risk of cancer.
Signal amplification
Figure 6: Signal amplification Pathways
Signal amplification is phenomenon in which when receptor proteins interact with the
signal molecules at the surface of the cell, in most cases signals are relayed to the
cytoplasm or the nucleus by second messengers which influences the activity of one or
more enzymes or genes inside the cell. However, most signalling molecules are found in
such a low concentration that their effect in cytoplasm would be minimal unless the
signal was amplified. Therefore, most enzymes linked and G-protein linked receptor use
a chain of other protein messenger to amplify the signal as it is being relayed. Thus in
case of protein kinase one cell surface receptor activates many G protein molecules. Each
G protein activates many adenylyl cyclases. Each cyclic AMP in turn will activate protein
kinase which then activates several molecules of a specific enzyme.
Figure 7: Signal amplification and cAMP
For example, the binding of a single molecule (such as glycogen or epinephrine) at the
cell surface can activate many effector G proteins and an adenylyl cyclase each of which
can produce a large number of cAMP messengers in a short period of time. Thus, the
production of a second messenger provides a mechanism to greatly amplify the signal
generated from the original message. There are many steps in the reaction cascade,
amplification of the signal via cAMP molecules which activate protein kinase K which
involve in phosphorylation of Ser, Thr and tyrosine of target protein. PKA is tertameric
protein which is made up of two catalytic and two regulatory subunits. Binding of cAMP
to the regulatory subunits induces a conformational change that leads to dissociation of
the catalytic subunits, which elicit formation of enzymatically active form of protein
kinase A, are now able to phosphorylate Ser and Thr residues on their target proteins. In
signal amplification, each PKA catalytic subunit phosphorylates a large number of
phosphorylase kinase molecules, which in turn phosphorylate an even larger number of
glycogen phosphorylase molecules, which in turn can catalyze the formation of a much
larger number of glucose phosphates. Thus, what begins as a hardly noticeable stimulus
at the cell surface is rapidly transformed into a major mobilization of glucose within the
cell.
Figure 8: Signal amplification Pathways for Glycogen degradation
The different effect of some hormone in different tissues in shown in the following table:
Table 1: Example of hormone induced response mediated by cAMP
Tissue Hormones Response

Liver Epinephrine and glucagon
Glycogen breakdown,
glucose synthesis
(glucogenesis), inhibition of
glycogen synthesis
Skeletal muscle Epinephrine Glycogen breakdown,
inhibition of glycogen
synthesis
Cardiac muscle Epinephrine Increase contractility
Adipose Epinephrine, ACTH and Triacyglycerol catabolism
glucagon
Kidney Vasopressin(ADH) Increase permeability of
epithelial cells to water
Thyroid TSH Secretion of thyroid
hormones
Bone Parathyroid hormone Increase calcium resorption
Ovary LH Increase secretion of steroid
hormones
Adrenal cortex ACTH Increase secretion of
glucocorticoids
Page 9 of 61
In the upcoming chapter we will study about each component of signal transduction in
detail.
Interesting Facts:
1. Most signaling molecules are found in such low concentration that their effect in
cytoplasm would be minimal unless the signal is amplified.
2. Most enzymes linked and G-protein linked receptor use a chain of other protein
messenger to amplify the signal as it being relayed.
Questions:
1. What is signal amplification?
2. What is Signal transduction? Explain diagrammatically.
3. Which are the possible factors which influence signal amplification?
4. How second messenger take part in the signal amplification pathway?
References:
1. Karp, G. Cell and molecular biology: concept and experiment. 6th edition,
chapter-15: Cell Signaling and Signal Transduction: Communication Between
Cells
2. http://www.biochem.mpg.de/en/eg/oesterhelt/web_page_list/ShortDesc_ST_casca
de/index.html
3. http://bcs.whfreeman.com/thelifewire/content/chp15/15020.html
4. Siso-Nadal, F., Fox, J.J., Laporte, S. A., Hebert, T. E., Swain, P.S. Cross-Talk
between Signaling Pathways Can Generate Robust Oscillations in Calcium and
cAMP. PLoS ONE, 2009, 4,
5. Stryer. Biochemistry, 5th edition.
M5 L2
Cell receptors
Receptor: A receptor is a protein molecule found on the surface of a cell which receives
chemical signals originating externally from the cell. Binding of specific signalling
molecules to a receptor directs a cell to allow certain molecules to enter or exit or directs
a cell to divide or die. Cells within multicellular organisms communicate via extracellular
mediators: either through diffusible molecules or by direct cell–cell contact. Examples of
receptors are G-Protein coupled receptors, Cytokine receptors as shown in Figure 1.
Receptors are located in either the cytoplasm or plasma membrane or nucleus of a cell. A
molecule which binds specifically to a receptor is called a ligand. A ligand may be
a peptide or other small molecules, such as a hormone, a neurotransmitter, a
pharmaceutical drug or a toxin. Each type of receptor recognizes and binds only certain
ligand shapes. Binding of a ligand to its receptor causes a conformational change in the
cytosolic domain of the receptor which then triggers the subsequent signalling cascade;
i.e. it activates or inhibits a specific biochemical pathway. Ligand-induced changes in
receptors result in cellular changes which constitute the biological activity of the ligands.
Most signalling molecules bind to receptors expressed on the target cell surface but some
signalling molecules are able to cross the plasma membrane and bind to intracellular
receptors in the cytoplasm or nucleus.
INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

G protein-coupled receptor Cytokine receptor
Receptor tyrosine kinases
Figure 1: Examples of receptors
Receptor types based on cellular location:

There are mainly three types of receptors present in the cell based on their location in the
cell. They are:
1. Cytosolic receptors
2. Nuclear receptors
3. Membrane bound receptors

1. Cytosolic receptors: Cytosolic receptors are specialized integral membrane proteins
that take part in communication between the cell and the outside world. Extracellular
signalling molecules (usually hormones, neurotransmitters, cytokines, growth factors or
cell recognition molecules) attach to the receptor, triggering changes in the function of
the cell. In this way the receptors play a unique and important role in cellular
communications and signal transduction. Most steroid hormones have receptors within
the cytoplasm which acts by stimulating the binding of their receptors to the promoter
region of steroid-responsive genes. Examples of cytosolic receptors are Estrogen
receptors, Glucocorticoid receptors etc.
Mechanism of action of cytosolic receptors: Estrogen diffuses across the plasma
membrane and binds to its receptor in the nucleus. The estrogen receptor is bound to Hsp
90 chaperones in the absence of the estrogen hormone as shown in Figure 2. The binding
of the estrogen induces a conformational change in the receptor, displacing Hsp 90 and
then leading to the formation of receptor dimers which binds to DNA, associate with
coactivators with histone acetyltransferase (HAT) activity, and stimulate transcription of
their target genes. In other cases, the receptor binds the DNA either in the presence or
absence of hormone. But hormone binding alters the activity of the receptor as a
transcriptional regulatory molecule. For example, in the absence of hormone, thyroid
hormone receptor is associated with a corepressor complex and represses transcription of
its target genes. Hormone binding induces a conformational change that results in the
interaction of the receptor with co activators rather than co repressors, leading to
transcriptional activation of thyroid hormone-inducible genes.
Estrogen Plasma membrane
Nucleus
Hsp 90 Receptor dimer

Coactivator
Inactive estrogen HAT

receptor
Acetyl group
Transcription
Figure 2: Estrogen receptor action

2. Nuclear receptors: Nuclear receptors are intracellular proteins expressed in the
nucleus of a cell. These receptors are members of a family of proteins known as the
nuclear receptor super family. Nuclear receptors constitute a superfamily of dimeric C4
zinc-finger transcription factors that bind lipid-soluble hormones and interact with
specific response elements in DNA. Steroid receptors are homodimers of zinc-finger
proteins that reside within the nucleus (except for the glucocorticoid receptor which
resides in the cytosol until it binds its ligand). Until their ligand finds them, some steroid
receptors within the nucleus associate with histone deacetylases (HDACs), keeping gene
expression repressed in those regions of the chromosome.
Structure of nuclear receptors: Nuclear receptors constitute a superfamily of dimeric

C4 zinc-finger transcription factors. They are modular in structure and contain the
following structural domains:
N-terminal regulatory domain (A-B): The A-B domain is highly variable in sequence
between various nuclear receptors. It contains the activation function 1 (AF-1) whose
action is independent of the presence of ligand. The transcriptional activation of AF-1 is
normally very weak but it synergizes with AF-2 in the E-domain to produce an
upregulation of gene expression.
DNA-binding domain; DBD (C): It is a highly conserved domain containing two zinc
fingers that binds to specific sequences of the DNA called hormone response elements
(HRE) as shown in Figure 3.
Hinge region (D): It is the flexible domain that connects the DBD with the LBD. It
influences subcellular distribution and intracellular trafficking.
Ligand binding domain LBD (E): Its sequence is moderately conserved but it is highly
conserved in structure between the various nuclear receptors. The structure of the LBD is
referred to as an alpha helical sandwich fold in which three anti parallel alpha helices (the
sandwich filling) are flanked by two alpha helices on one side and three on the other (the
bread). The ligand binding cavity is within the interior of the LBD and just below is
present three anti parallel alpha helical sandwich filling. Along with the DBD, the LBD

contributes to the dimerization interface of the receptor. In addition, it binds coactivator
and corepressor proteins. The LBD also contains the activation function 2 (AF-2) whose
action is dependent on the presence of bound ligand.
C-terminal domain (F): It is highly variable in sequence between various nuclear

receptors.
Figure 3: Structural Organization of Nuclear Receptors (estrogen receptor)

Top – Schematic amino acid sequence of a nuclear receptor.
Bottom – 3D structures of the DBD (bound to DNA) and LBD (bound to hormone) regions of the nuclear receptor
Mechanism of action: The intracellular nuclear receptors respond to small hydrophobic

signaling molecules that diffuse readily across the plasma membrane. These molecules
bind to the receptors and a conformational change takes place in the receptor. This is
followed by a series of intracellular signal transduction cascade. The steroid hormones
like thyroid hormone, vitamin D3 and retinoic acid differ greatly from one another in both
chemical structure and function. Once inside the cell, these signaling molecules bind to
intracellular receptors that are expressed by the hormonally responsive target cells. These
receptors are transcription factors that contain domains for ligand binding, DNA binding
and transcriptional activation. Ligand binding regulates their function as activators or
repressors of their target genes. So the steroid hormones and related molecules directly
regulate gene expression. In response, these receptors work with other proteins to
regulate the expression of specific genes, thereby controlling the development,
homeostasis, and metabolism of the organism.

The nuclear receptor is kept in the cytoplasm by interaction between its ligand-binding
domain (LBD) and inhibitor proteins in the absence of a steroid hormone. When hormone
is present, it diffuses readily through the plasma membrane and binds to the ligand-
binding domain. This causes a conformational change thus releasing the receptor from
the inhibitor proteins. The receptor with bound ligand is then translocated into the
nucleus, where its DNA-binding domain (DBD) binds to response elements, allowing the
ligand-binding domain and an additional activation domain (AD) at the N-terminus to
stimulate transcription of target genes as shown in Figure 4.
Figure 4: Model of hormone-dependent gene activation by a homodimeric nuclear receptor.
3. Membrane bound receptors: Membrane bound receptors are proteins that are
associated with the cell membrane. They can span across the membrane and can transmit
a signal from outside the cell to inside the cell. Outside the cell, a ligand (e.g. Hormone)
will bind to the receptor. A few chemical stimuli, including steroid hormones and the gas
nitric oxide cross the plasma membrane and bind receptors inside the cell. Thus the
receptor undergoes a conformational change. This change in the shape of the receptor is
detected inside the cell. It is the shape change that is the transmission of the signal from
the outside to the inside. Inside the cell, other proteins can interact with the receptor in its
new shape and be turned 'on' to continue the signal pathway.

Selective expression of certain receptors and their associated cytoplasmic transduction
machinery allows differentiated cells to respond specifically to particular ligands.
Members of each family of receptors share one or more structurally homologous
domains. In some families, the members share both ligand binding and signal transducing
strategies (seven helix receptors, G-protein coupled receptors, cytokine receptors).
Members of other families share either a similar ligand-binding structure (Tumour
Necrosis Factor receptor family) or a common signal transducing method (receptor
tyrosin kinases).
Membrane receptors are found also in the cis Golgi network which captures the proteins
during protein sorting and carries them in transport vesicles back to the ER. Also most
cholesterol is transported in the blood bound to protein in the form of particles known as
low density lipoproteins or LDL. When a cell needs cholesterol for membrane synthesis,
it makes transmembrane receptor for LDL and inserts them into its plasma membrane.
Once in the plasma membrane, the LDL receptors diffuse until they associate with
clathrin-coated pits as shown in Figure 5.
Figure 5: LDL receptor proteins binding to a coated pit in the plasma membrane of a cell

The seven major classes of cell surface receptors are:
1. G protein-coupled receptors
2. Cytokine receptors
3. Receptor tyrosine kinases
4. TGFβ receptors
5. Hedgehog receptors
6. Wnt receptors
7. Notch receptor
Structure of Seven helix receptors: Membrane bound receptors constitute the members
of the largest family of plasma membrane receptors built from a serpentine arrangement
of seven transmembrane α helices. G protein–coupled receptors (GPCRs) are examples of
seven transmembrane α helices receptors. GPCRs are found in all eukaryotic cells from
yeast to man. All G protein–coupled receptors contain seven membrane-spanning regions
with their N-terminal segment on the exoplasmic face and their C-terminal segment on
the cytosolic face of the plasma membrane.

As shown in Figure 6 (a), all receptors of this type have the same orientation in the
membrane and contain seven transmembrane - helical regions (H1–H7) - four
extracellular segments (E1–E4), and four cytosolic segments (C1–C4). The carboxyl-
terminal segment (C4), the C3 loops and, in some receptors, also the C2 loops are
involved in interactions with a coupled trimeric G protein. The long C3 loop between
helices 5 and 6 is important for interactions between a receptor and its coupled G protein.
This superfamily of seven-pass transmembrane receptor proteins includes rhodopsin, the
light-activated protein in the vertebrate eye as shown in Figure 6 (b).
Figure 6 (a): Schematic diagram of G protein–coupled receptors
Figure 6 (b): Atomic structure of seven helix receptor of bovine rhodopsin, the light activated protein in vertebrate eye

Mechanism of action of seven helix receptors: The seven helix receptors use trimeric
GTP-binding proteins to relay signals to effector proteins inside the cells. Human genome
encodes several thousand GPCR. These include receptors in the visual, olfactory (smell)
and gustatory (taste) systems, neurotransmitter receptors and most of the receptors for
hormones that control the metabolism of carbohydrate, amino acid and fat. GPCRs are
coupled to signal-transducing trimeric G proteins. In mammals, olfactory cells alone use
500 – 1000 different seven-helix receptors to discriminate odorant molecules.
Phosphorylation of the C-terminal tail inactivates many types of seven helix receptors.
Two different strategies, sometimes acting on the same receptor, provide negative
feedback. One strategy is for the second messengers, which are produced in response to
receptor activation to stimulate general protein kinases (including cAMP), protein kinase
A (PKA) and protein kinase C (PKC) which phosphorylate the activated receptor.
Phosphorylation inhibits the receptor thus allowing for crosstalk between receptors. The
second strategy involves a class of protein kinases specific for the receptor themselves.
They are called G-protein coupled receptor kinases. These kinases phosphorylate
multiple serines or threonines on the C-terminal cytoplasmic tail of active receptors.
Phosphorylation promotes binding of a regulatory protein called arrestin which
inactivates the receptor by blocking interaction of the receptor with trimeric G-proteins.
Arrestin binding to some seven helix receptors promotes their removal from the plasma
membrane by endocytosis. G protein–coupled receptors transduce signals from
extracellular hormones to associated effector proteins. In the resting state, when no ligand
is bound to the receptor, the Gα subunit is bound to GDP and complexed with Gβγ. As
shown in the Figure 6, ligand binding shifts the equilibrium from the resting
conformation towards the active conformation. Active receptor promotes dissociation of
GDP from α subunit of multiple trimeric G-proteins, allowing GTP to bind. This
dissociates Gα from Gβγ, each of which activate downstream effectors that produce the
second messengers cAMP and diacylglycerol (DAG) as shown in Figure 7. cAMP and
DAG activates PKA and PKC, which phosphorylate active receptors on their C-terminus.
This attracts arrestin, putting the receptor into the inactive adapted state.

Figure 7: Activation and adaptation of a seven helix receptor: A, resting; B, active; C, adapted.
Epinephrine Binds to Several Different G Protein–Coupled Receptors. All epinephrine

receptors are G protein– coupled receptors, the different types are coupled to different G
proteins. These receptors are of interest because they trigger different intracellular signal-
transduction pathways. Both subtypes of β-adrenergic receptors, termed β1 and β2, are
coupled to a stimulatory G protein (Gs) that activates the membrane-bound enzyme
adenylyl cyclase. Once activated, adenylyl cyclase catalyzes synthesis of the second
messenger cAMP. That binding of epinephrine to β-adrenergic receptors induces a rise in
cAMP.
Interesting facts:
• Receptor tyrosine kinase is the first receptor to be discovered.
• G proteins were discovered by Alfred G. Gilman and Martin Rodbell when
investigating stimulation of cells by adrenaline.
• Skin contains approximately 640,000 sense receptors scattered unevenly over the
body’s surface.
Questions:
1. What are receptors? Give examples.
2. How many types of receptors are present on the cell?
3. What are cytosolic receptors? Give example.
4. What are nuclear receptors?
5. What is membrane bound receptors? Give example.

References:
Alberts, B., Johnson, A., Lewis, J., Raff, M., Roberts, K. (2002). Molecular Biology of
the Cell, 4th ed. Taylor & Francis, Inc.
Cooper, G. M., Hausman, R. E. (2007). The Cell: A Molecular Approach, 4th ed. Sinauer
Associates, Inc.
Lodish, H., Berk, A., Matsudaira, P., Kaiser, C. A., Kreiger, M., Scott, M. P., Zipursky,
L., Darnell, J. (2003). Molecular Cell Biology, 5th ed. Freeman, W. H. & Company.

M5 L3
Second messengers - cAMP
Second messengers: As mentioned din lecture 1 of this module, second messengers are
molecules that relay signals received at receptors on the cell surface such as hormones,
growth factors, etc. to appropriate target molecules in the cytosol and/or nucleus. In
addition to their job as relay molecules, second messengers serve to amplify the strength
of the signal. Binding of a ligand to a single receptor at the cell surface may end up
causing massive changes in the biochemical activities within the cell.
There are 3 major classes of second messengers:
1. Cyclic nucleotides (cAMP and cGMP)

2. Inositol trisphosphate (IP3) and diacylglycerol (DAG)
3. Calcium ions (Ca2+)
We will discuss about all of them in the upcoming lectures.
Cyclic adenosine monophosphate (cAMP)

cAMP is an important second messenger involved in a plethora of cellular effects and
biological roles by regulating various metabolic process and mediating the effects of
many hormones that binds to a specific receptor on the cell membrane of target cells
including catecholamines, ACTH, and vasopressin. It also plays imperative role in the
transcription of some genes. Earl Sutherland won a Nobel Prize in Physiology or
Medicine in 1971 for his discoveries regarding the mechanisms of the action of
hormones, especially epinephrine, via second messengers such as cyclic AMP. cAMP is
represented by C10H12N5O6P and the molecular mass is 329.206.
NH2
N
N
CH2 N
O N
H H
O
H H
HO P
O OH
O
Figure 1: Cyclic Adenosine monophosphate or 3'-5'-cyclic adenosine monophosphate

[Compositions of cAMP - Adenine base + Ribose sugar + 3’, 5’-cyclic phosphate]
Adenosine in cAMP is a nucleoside composed of the pentose sugar D-ribose and adenine,
a base. Cyclic AMP contains an ester linkage between the phosphate and ribose units.
Some of the hormones that achieve their effects through cAMP as a second messenger:
• Adrenaline
• Glucagon
• Luteinizing hormone (LH)
Binding of the hormone to its receptor activates a G protein which, in turn, activates
adenylyl cyclase. The resulting rise in cAMP turns on the appropriate response in the cell
by either (or both): changing the molecular activities in the cytosol, often using Protein
Kinase A (PKA) — a cAMP-dependent protein kinase that phosphorylates target
proteins; turning on a new pattern of gene transcription.
Functions of cAMP
1. cAMP as a second messenger: cAMP is a second messenger, used for intracellular
signal transduction. It is involved in transmitting signal from outside the cell to the
interior via the process of binding of hormones like glucagon and epinephrine or other
signal molecules to cell membrane receptor. It is involved in the activation of protein
kinases and regulates the effects of adrenaline and glucagon. cAMP also binds to and
regulates the function of ion channels such as the HCN channels and a few other cyclic
nucleotide-binding proteins such as Epac1.

2. Regulation of protein kinase A by cAMP (Phosphorylation of protein kinase): The
most important function of cAMP in animal cell is regulation of protein kinase A activity.
Protein kinase A is found primarily in inactive form in the cell in which it consists of two
regulatory (R) and two catalytic (C) subunits together. Binding of cAMP to the regulatory
subunits induces a conformational change that leads to dissociation of the catalytic
subunits, which elicits formation of enzymatically active form of protein kinase A, and
are now able to phosphorylate Ser and Thr residues on their target proteins.
Figure 2: Regulation of protein kinase A

3. Regulation of Cyclic AMP-inducible gene expression via protein kinase A:
In many animal cells, increase in cAMP activates the transcription of specific target
genes that contain a regulatory sequence called the cAMP response element, or CRE. In
this case, the signal is passed from the cytoplasm to the nucleus by the catalytic subunit
of protein kinase A, which is able to enter the nucleus after its release from the regulatory
subunit. Within the nucleus, protein kinase A phosphorylates a transcription factor called
CREB (CRE-binding protein), leading to the activation of cAMP-inducible genes. Thus
such type of regulation of gene expression by cAMP plays significant role in controlling
proliferation, survival, and differentiation of a wide variety of animal cells.
Figure 3: Cyclic AMP inducible gene expression
4. In eukaryotic cells: cAMP and its associated kinases play key role in numerous
biochemical processes, including the regulation of glycogen, sugar, and lipid metabolism.
There are several minor PKA-independent functions of cAMP like activation of calcium
channels, providing a minor pathway by which growth hormone-releasing hormone
causes a release of growth hormone. The GEF (guanine nucleotide exchange factor)

domain is usually covered by the N-terminal region containing the cAMP binding
domain. When cAMP binds, the domain dissociates and exposes the active GEF domain,
allowing Epac to activate small Ras-like GTPase proteins, such as Rap1.
5. In bacteria: In bacteria, cAMP plays a crucial role and its level varies depending on
the medium used for growth. In E.coli, cAMP involves in the positive regulation of the
lac-operon. cAMP is synthesized from ATP by adenylyl cyclase, as a result increase in
the level of cAMP causes decrease in glucose concentration which is the carbon source.
cAMP then binds to the transcriptional regulatory protein, cAMP receptor protein (CRP)
also called catabolic activator protein(CAP). After binding of cAMP to CAP enhance the
binding capacity of CAP to its binding site (CAP binding site) on target DNA sequence
which in lac operon located 60 nucleotides upstream of transcription start site, making it
easier for RNA polymerase to bind to the adjacent promoter to start transcription of the
lac-operon, increasing the rate of lac-operon transcription. With a high glucose
concentration, the cAMP concentration decreases, and the CRP disengages from the lac-
operon.
Figure 4: Positive regulation of lac operon by glucose repression coupled to enhance level of cAMP.

6. In some slime moulds: In some slime mold species such as Dictyostelium discoideum,
the chemotactic movement of cells is organized by periodic waves of cAMP that
propagates through the cell. The waves are the result of a regulated production and
secretion of extracellular cAMP and a spontaneous biological oscillator that initiates the
waves at centers of territories.
7. Mitochondrial Biogenesis: cAMP/PKA has key role in mitochondrial compartment

biogenesis. Mitochondria takes part in several vital cellular functions. For example they
are involved in ATP production, via the process of oxidative phosphorylation, the ATP
production via mitrochondria is 15 times more than glycolysis alone. They also take
central part in metabolic regulation and assist diverse cell signalling events. Mitochondria
are therefore essential for the maintenance, adaptability and survival of eukaryotic cells.
cAMP/PKA signaling balances respiratory activity with mitochondria dependent
apoptosis via transcriptional regulation.
Areas of action of cyclic AMP:
There are so many application of cAMP for the prevention of several disorders in human
beings. Some are shown below:

Positive
Inotropic
Action in Heart
Prevent
Anti Glaucoma platelet
Aggregation
cAMP
Reduce Blood Bronchodilation
Pressure
Reduce
Inflammatiom
Figure 5: Action of cAMP in our body
Mechanism of Regulation of cAMP: When stimulatory hormone binds to G-protein

coupled receptor some conformational changes occur in the receptor by which its
unexposed catalytic part gets exposed due to which it can interact and activate G-protein
with exchange of GDP by GTP at Gα subunit. Now Gα subunit goes and activates
membrane bound adenylyl cyclase that facilitates the conversion of ATP to cAMP. This
cAMP is now ready to activate protein kinase A which is involved in phosphorylation of
several proteins that takes part in the further cellular responses. At the same time of
cAMP production by adenylyl cyclase, the cAMP level is controlled by other enzyme that
is called cAMP phosphodiesterase which converts cAMP into 5’-AMP. On the other
hand, when inhibitory hormone binds to G-protein coupled receptor, it counters the effect
as shown in Figure 6. In this process, the adenylyl cyclase is inhibited because GPCR
activates the G protein that contains inhibitory alpha subunit which binds to enzyme and
inhibits cAMP production.

Figure 6: Synthesis and Regulation of cAMP
Synthesis and Degradation of cAMP: cAMP is a cyclic nucleotide which serves as an

intracellular and in some cases extracellular secondary messenger. It is derived from
adenosine triphosphate (ATP) by adenylyl cyclase located on inner side of plasma
membrane and used for intracellular signal transduction in many different organisms,
conveying the cAMP-dependent pathway. The elevated cAMP level is regulated by
degradation pathway which takes place by cAMP phosphodiesterase enzyme.
O
O -
- H2 O P O CH2 O Adenine
O P O CH2 O Adenine
C O Adenine O- H H
O H H H
H
-
O P O H H H H
Aden ylyl O cAMP
cyclase H H
O - Phosphodiesterase OH OH
OH OH O P
O OH
- P
O O O
ATP cAMP 5' AMP
O-
Figure 7: Synthesis and degradation of cAMP
Interesting facts:
1. Cyclic AMP is synthesized from ATP by the action of the enzyme adenylyl cyclase.
2. cAMP decomposition into AMP is catalyzed by the enzyme phosphodiesterase.

3. cAMP and its associated kinases function in several biochemical processes, including
the regulation of glycogen, sugar, and lipid metabolism.
Questions:
1. Explain the effect of glucose concentration on the cAMP and catabolic activator
protein?
2. How cAMP involve in signal transduction pathway?
3. How cAMP synthesizes and regulated in our body?
References:
1. Horton, R. A., Moran, L. A., Scrimgeour, G., Perry, M. and Rawn, D. Principle of
biochemistry. 6th edition 2006 pearson prenticle.
[http://sandwalk.blogspot.in/2007/05/regulating-glycogen-metabolism.html]
2. Cooper, G.M. The Cell: A Molecular Approach. fouth edition.
3. http://en.wikipedia.org/wiki/Cyclic_adenosine_monophosphate
4. Yoboue, E. D., Augier, E., Galinier, A., Blancard, C., Pinson, B., Casteilla, L.,
Rigoulet, M. and Devin, A. cAMP-induced mitochondrial compartment
biogenesis: role of the glutathione redox state
5. Leadsham J. E. and Gourlay, C. W. cAMP/PKA signaling balances respiratory
activity with mitochondria dependent apoptosis via transcriptional regulation.
BMC cell biology, 2010

M5 L4
Second messengers - cGMP
Cyclic Guanosine monophosphate: Cyclic Guanosine monophosphate or cGMP, is a
cyclic nucleotide derived from guanosine triphosphate(GTP). cGMP is a multi-functional
second messenger molecule, similar in action to cAMP but generally producing opposite
effects on cell function. It has molecular formula of C10H12N5O7P and has molecular
weight of 345.2 g/mol. It has composition of Guanine nucleotide base, ribose sugar and
cyclic phosphate between 3’ and 5’ positions of ribose sugar as shown in figure 1. Cyclic
GMP is synthesized from the nucleotide GTP using the enzyme guanylyl cyclise as
shown in figure 2.
Structure:
Figure 1: Guanosine 3',5'-cyclic phosphate
Synthesis of cGMP: cGMP is an important signal molecule that carries different

messeges in different tissues. cGMP is generated via two pathways distinguished by the
nature of Guanylyl Cyclase (GC) that mediate its conversion from guanosine triphosphate
(GTP). The guanyl cyclase is generally found in cell in two forms - soluble form and
membrane- bound form. They are generated via two pathways as described below:
1. The soluble pathway, where cGMP is generated via nitric oxide (NO)-activated
guanylyl cyclase which is cytosolic protein with tightly associated heme group. NO is
sufficiently non-polar which can easily cross the plasma membrane of target cell without
any carrier and binds to the heme group of guanylyl cyclase and activates the cGMP
production.

2. In the membrane-bound pathway, GCs is transmembrane protein with extracellular
ligand binding domain, share some homology with those activated by NO. The ligands
for a subset of membrane GCs are members of the Natriuretic Peptide (NP) hormone
family including atrial NP hormone, B-type NP hormone and C-type NP hormone.
Figure 2: Conversion of GTP to cGMP
Functions of cGMP:
1. cGMP is an important molecule of the cell that takes part in various activities in
cellular system. When guanylyl cyclase stimulation leads to elevated levels of cGMP, it
then mediates biological responses, such as blood vessel dilation which increases blood
flow.
2. The action of cGMP is regularly facilitated by stimulation of cGMP dependent protein
kinases, although cGMP is a common regulator of ion channel conductance,
glycogenolysis, cellular apoptosis and phosphodiesterases.
3. Another well-known role of cGMP is in the vertebrate eye, where it serves as the
second messenger responsible for converting the visual signals received as light to nerve
impulses. The photoreceptor in rod cells of the retina is a G protein-coupled receptor
called rhodopsin. When light falls on the extracellular side of rhodopsin, then some
conformational changes occurs in it by which its bounded chromophore 11-cis retinal is
converted to all-trans retinal form, ultimately rhodopsine’s unexposed catalytic
cytoplasmic side gets exposed which interacts with the G protein transducin and activates
them by replacement of GDP by GTP on its α- subunit. The activated Gα then activates
cGMP phosphodiesterase 6 which converts all cGMP into 5’ GMP. Due to this cGMP
level gradually decreases, the cGMP dependent sodium ion-channel becomes closed. This

channel is also entry site of calicium ions so Ca++ level also decrease. This critical
situation created in the cell is called hyperpolyrisation. But after decreasing Ca++ level,
guanylyl cyclase is activated and again cGMP synthesis starts. Thus we can summarize
the whole phenomenon as, change in cGMP level in retinal rod cells is translated to a
nerve impulse by a direct effect of cGMP on ion channels in the plasma membrane.
Figure 3: Visual Signal Transduction. The light-induced activation of rhodopsin leads to the hydrolysis of cGMP, which in
turn leads to ion channel closing and the initiation of an action potential.
4. In kidney, membrane bound guanylyl cyclase is activated by the hormone atrial

natriuretic factor (ANF), which is released by cells in atrium of the heart when the heart
is stretched by increased blood volume. ANF comes to kidney with blood from heart
and activates membrane bound guanylyl cyclase in the cells of collecting ducts. The
resulting rise in cGMP level in cells triggers increase renal excretion of Na+ and water,
driven by change in osmotic pressure. Water loss causes reduction of blood volume
which counters the production of ANF. Vascular smooth muscle also contains ANF
receptor-guanylyl cyclase which release. On binding to these receptor , ANF causes
relaxation of blood vessels which increases the blood flow with decreasing blood
pressure.
cGMP, like cAMP get synthesized by receiving odourous input by olfactory receptor.
cGMP is produced slowly and has a more sustained life than cAMP. cGMP in the
olfactory is synthesized by both membrane guanylylcylcase (mGC) as well as soluble
guanylyl cyclase (sGC). cGMP synthesis in the olfactory is due to sGC activation by
nitric oxide, a neurotransmitter. cGMP also requires increased intracellular levels of

cAMP and the cross-link between the two second messengers seems to be due to rising
intracellular calcium levels.
Vasodilation: NO is an extracellular gaseous second messenger. NO is unusual because
it acts both as an extracellular messenger, mediating intercellular communication, and
as a second messenger, acting within the cell in which it is generated. NO is
synthesized by L-arginine which is catalyzed by nitric oxide synthesae. The NO
formed in the endothelial cell diffuses across the plasma membrane and into the
adjacent smooth muscle cells, where it binds and stimulates guanylyl cyclase which
synthesizes cyclic GMP (cGMP).
Figure 4: Signal transduction pathway leads to dilation of blood vessel through NO and cGMP.

Figure 5: Pathways of generation of cGMP and several areas of effect
The conversion mechanism of GTP to cGMP by guanyl cyclase is shown below:
Regulation Pathway of cGMP: In animals, most of the actions of cGMP are supposed
to be mediated by cGMP-dependent protein kinase, which is also called protein kinase G,
abbreviated as PKG. After activation of PKG by cGMP, phosphorylates Ser and Thr
residues in target proteins. PKG has catalytic and regulatory domains on a single
polypeptide chain (Mr, 80,000). A part of regulatory domains fit comfortably in
substrate-binding site. After binding of cGMP forces this part of regulatory domain out of
the binding site, activating the catalytic domain.

Figure 6: Regulatory Pathway of cGMP
Interesting facts:
1. Cyclic GMP is synthesized from the nucleotide GTP using the enzyme guanylyl
cyclase.
2. Cyclic GMP serves as the second messenger for atrial natriuretic peptide (ANP),
nitric oxide (NO), the response of the rods of the retina to light.
3. Some of the effects of cGMP are mediated through Protein Kinase G (PKG) a
cGMP-dependent protein kinase that phosphorylates target proteins in the cell.
Questions:
1. How cGMP is involved in vasodilation or smooth muscle relaxation?
2. What is the visual pathway for light transduction in brain?
3. How cGMP regulation take place and what are the enzyme involve in its
regulation?

References:
1. http://medical-
dictionary.thefreedictionary.com/cyclic+guanosine+monophosphate
2. http://www.nature.com/embor/journal/v7/n2/fig_tab/7400627_f1.html
3. http://www.sigmaaldrich.com/life-science/cell-biology/learning-center/pathway-
slides-and/cyclic-nucleotide-metabolism-cgmp.html
4. Berg, J., Tymoczko, J. and Stryer, L. Biochemistry. 5th edition, chapter-15
5. Wikipedia
6. Nisoli, E., Cozzi, V., Carruba, M. O. Amino acids and mitochondrial biogenesis.
The American Journal of Cardiology,2008, Volume: 101, Issue: 11A, Pages:
22E-25E
Cells

M5 L5
Calcium ion flux and its role in cell signalling
Calcium ion flux: Calcium ions are also important intracellular messengers. In fact,
calcium ions are probably the most widely used intracellular messengers. Calcium (Ca2+)
plays an essential role in the physiology and biochemistry of organisms and the cell. It
plays role in common signalling mechanism because once it enters the cytoplasm it exerts
allosteric regulatory affects on many enzymes and proteins. Calcium is a second
messenger produced by indirect signal transduction pathways such as G-protein coupled
receptors. Calcium ions (Ca2+) impact nearly every aspect of cellular life. The principles
of Ca2+ signaling, from changes in protein conformations driven by Ca2+ to the
mechanisms that control Ca2+ levels in the cytoplasm and organelles. The highly
localized nature of Ca2+-mediated signal transduction and its specific roles in excitability,
exocytosis, motility, apoptosis, and transcription. Normally, cytosolic calcium [Ca2+] ions
is kept very low (10-7 M) by the action of Ca2+ pumps in the ER, mitochondria and
plasma membrane. Hormonal, neural, or other stimuli cause either an influx of Ca2+ into
the cell through specific Ca2+ channels in the plasma membrane or the release of
sequestered Ca2+ from the ER or mitochondria, in either case raising the cytosolic [Ca2+]
and triggering a cellular response. This phenomenon is called Calcium ion flux.
Role of Calcium in cell signalling:

In response to many different signals, a rise in the concentration of Ca2+ in the cytosol
triggers many types of events such as:
• Muscle contraction
• Exocytosis
a) Release of neurotransmitters at synapses (and essential for the long-term synaptic
changes that produce Long-Term Potentiation (LTP) and Long-Term Depression
(LTD)
b) Secretion of hormones like insulin
• Activation of T cells and B cells when they bind antigen with their antigen receptors
(TCRs and BCRs respectively)

• Adhesion of cells to the extracellular matrix (ECM)
• Apoptosis
• A variety of biochemical changes mediated by Protein Kinase C (PKC)
The concentration of calcium ions in a particular cellular compartment is guarded by the

regulated activity of Ca2+ pumps, Ca2+ exchangers, and/or Ca2+ ion channels located
within the membranes that surrounds the compartment. There are two major types of
signaling receptors- G-protein coupled receptor (GPCRs) and receptor protein-tyrosine
kinases (RTKs). One of the most important pathways of intracellular signalling is based
on the use of second messengers derived from the membrane phospholipids
phosphatidylinositol 4, 5-bisphosphate (PIP2). PIP2 is a minor component of the plasma
membrane, localized to the inner leaflet of the phospholipids bilayer. Hydrolysis of PIP2
by phospholipase C-β is stimulated by variety of hormones and neurotransmitters. After
hydrolysis PIP2 is cleaved into two components- Diacyl glycerol (DAG) and inositol
l,4,5- triphosphate (IP3), both are also the secondary messengers of cell. Diacylglycerol
and IP3 stimulate distinct down-stream signalling pathways (protein kinase C and Ca2+
mobilization, respectively), so PIP2 hydrolysis triggers a two-armed cascade of
intracellular signalling.
The first secondary messenger diacylglycerol produced by hydrolysis of PIP2 remains
associated with the plasma membrane and activates protein-serine/threonine kinases
belonging to the protein kinase C family, many of which play important roles in the
control of cell growth and differentiation.

The other second messenger produced by PIP2 cleavage, IP3, is a small polar molecule
that is released into the cytosol, where it acts to signal the release of Ca2+ from
intracellular stores.
Figure 1: Ca2+ mobilization by IP3

Whereas the other important pathway in which extracellular messengers that signal
through RTKs can trigger a similar response as in GPCR signal pathway. The major
difference is that RTKs activate members of the phospholipase C-γ subfamily, which
possess an SH2 domain that allows them to bind to the activated, phosphorylated RTK.
Figure 2: Comparative pathway of GPCR and RTKs for release of calcium from ER
There are numerous other PLC isoforms. For example, PLC δ is activated by Ca2+ ions,
and PLCε is activated by Ras-GTP. All PLC isoforms carry out the same reaction,

producing IP3 and linking a multitude of cell surface receptors to an increase in
cytoplasmic Ca2+. There is another major route leading to elevation of cytosolic [Ca2+]
which involve in synaptic transmission. In this case, a nerve impulse leads to a
depolarization of the plasma membrane, which triggers the opening of voltage-gated
calcium channels in the plasma membrane, allowing the influx of Ca2+ ions from the
extracellular medium.
Properties of Calcium Ion (Ca2+)
There are two major properties that allow Calcium (Ca2+) ion to work effectively as a
signaling mechanism:
 Ca2+ levels inside the cell are readily detectable. This is because the levels of Ca2+ are
highly regulated by transport systems that expel Ca2+ from the cell. The level of Ca2+
in the cytoplasm is approximately 100nM, which are several orders of magnitude
lower than outside the cell as a result of Ca2+ pumps that actively export Ca2+ from the
cell. Ca2+ is pumped not only across the plasma membrane but also into the
endoplasmic reticulum, which therefore serves as an intracellular Ca2+ store. IP3 acts to
release Ca2+ from the endoplasmic reticulum by binding to receptors that are ligand-
gated Ca2+ channels. As a result, cytosolic Ca2+ levels increase to about 1 μM, which
affects the activities of a variety of target proteins, including protein kinases and
phosphatases.
• Ca2+ can readily bind to proteins and cause conformational changes. Ca2+ is attracted
to the negatively charged oxygen atoms in the side chains of glutamate and
asparagines, and the uncharged oxygen in both the side chains and main chains of
glutamine and asparagine. Ca2+ is readily able to cause large conformational changes
due to the fact that it can form ligand with up to eight oxygen atoms. This can lead to
cross linking of amino acids in a protein that did not exist before Ca2+ was introduced.
Function of Ca2+ in cell:

• Protein function is governed by shape and charge. Ca2+ binding triggers changes in
protein shape and charge. Similarly, phosphorylation imparts a negative charge,
altering protein conformations and their interactions. Protein kinases, comprising
~2% of eukaryotic genomes, remove phosphate from ATP and covalently attach it to
the free hydroxyl groups of serine, threonine, or tyrosine residues. The abilities of
Ca2+ and phosphate ions to alter local electrostatic fields and protein conformations
are the two universal tools of signal transduction.
• Ca2+ had a new and totally unexpected function: it carried the information necessary
for the contraction of heart.
 Changes in intracellular [Ca2+] are detected by Ca2+-binding proteins that regulate a
variety of Ca2+-dependent enzymes.
Example of Ca2+-dependent enzymes:
o Calmodulin (CaM) (Mr 17,000) is an acidic protein with four high-affinity Ca2+-
binding sites. When intracellular [Ca2+] rises to about 10-6 M (1 μM), the binding of
Ca2+ to calmodulin drives a conformational change in the protein. Calmodulin
associates with a variety of proteins and, in its Ca2+-bound state, modulates their
activities. Calmodulin is a member of a family of Ca2+-binding proteins that also
includes troponin, which triggers skeletal muscle contraction in response to increased
[Ca2+]. This family shares a characteristic Ca2+-binding structure.
o Calmodulin is also an integral subunit of a family of enzymes, the Ca2+/calmodulin-
dependent protein kinases (CaM kinases I–IV). When intracellular [Ca2+] increases in
response to some stimulus, calmodulin binds Ca2+, undergoes a change in
conformation, and activates the CaM kinase. The kinase then phosphorylates a number
of target enzymes, regulating their activities. Calmodulin is also a regulatory subunit
of phosphorylase b kinase of muscle, which is activated by Ca2+. Thus Ca2+
triggers
io
ATP-requiring muscle contract ns while also activating glycogen breakdown,
providing fuel for ATP synthesis. Many other enzymes are also known to be
modulated by Ca2+ through calmodulin.

Figure 3: Calcium ion dependent enzyme- calmodulin and Ca2+/calmodulin-
dependent protein kinases
• The rise of intracellular free Ca2+ ([Ca2+]i) is an crucial triggering signal for T-cell
activation by antigen and other stimuli that cross-link the T cell antigen receptor (TCR).
• The downstream consequences of Ca2+ signalling, also involve in gene expression on
the influence of signal complexity.

Some Proteins Regulated by Ca2+ and Calmodulin:
• Adenylyl cyclase (brain)

• Ca2+/calmodulin-dependent protein kinases (CaM kinases I to IV)
• Ca2+-dependent Na+ channel (Paramecium)
• Ca2+_-release channel of sarcoplasmic reticulum
• Calcineurin (phosphoprotein phosphatase 2B)
• cAMP phosphodiesterase
• cAMP-gated olfactory channel
• cGMP-gated Na+, Ca2+ channels (rod and cone cells)
• Glutamate decarboxylase
• Myosin light chain kinases
• NAD+ kinase
• Nitric oxide synthase
• Phosphoinositide 3-kinase
• Plasma membrane Ca2+ ATPase (Ca2+ pump)
• RNA helicase (p68)
Interesting facts:
1. Calcium ions are probably the most widely used intracellular messengers.
2. Normally, the level of calcium in the cell is very low (~100 nM).
3. Getting Ca2+ into (and out of) the cytosol is via Voltage-gated channels.
Questions:
1. Ca2+, IP3 and cAMP have all been described as second messengers. In what ways
are their mechanisms of action similar? In what ways are they different?
2. Describe the function of Ca2+ as a second messenger.
References:
1. Cooper, G.M. The Cell: A Molecular Approach. fouth edition. Chapter-15: Cell
Signaling.
2. Karp, G. “Cell and Molecular Biology-Concept of Experiment” 6th edition.
Chapter-15: Cell Signaling and Signal Transduction: Communication Between
Cells.

M5 L6
G proteins in signal transduction
G- Protein: G proteins (guanine nucleotide-binding proteins) are a family of proteins
involved in transmitting chemical signals outside the cell, and causing changes inside the
cell. They communicate signals from many hormones, neurotransmitters, and other
signaling factors.
Type of G protein:
G protein can refer to two distinct families of proteins.
• Heterotrimeric G proteins: sometimes also known as the large G proteins that are
activated by G protein-coupled receptors and made up of alpha (α), beta (β), and gamma
(γ) subunits.
• Small G proteins: They are proteins of 20-25kDa that belong to the Ras superfamily
of small GTPases. These proteins are homologous to the alpha (α) subunit found in
heterotrimers, and are in fact monomeric. However, they also bind GTP and GDP and are
involved in signal transduction.
Heterotrimeric G-protein
Heterotrimeric G proteins are more complex protein which were first characterized by
Martin Rodbellare. It consists of three different subunits- α, β, and γ having molecular
weight of these are 45, 37, and 9 kD respectively, among these α subunit binds to GDP in
unactive state or GTP in active state, hence heterotrimeric G-protein is known as a
member of the G protein superfamily. Heterotrimeric G proteins are held at the plasma
membrane by lipid chains that are covalently attached to the α and γ subunits.
Structure of Heterotrimeric G-protein:
Figure 1: Structure of Heterotrimeric G-protein

The alpha subunit has two domains - the transducing insertion domain fold whereas the
other is a P-loop containing nucleoside triphosphate hydrolase fold. P-loop or a
phosphate-binding loop is an ATP or GTP- binding site motif found in many nucleotide-
binding proteins. It is a glycine-rich loop led by a beta sheet and followed by an alpha
helix. It interacts with the nucleotide phosphate groups and with the Mg2+ ion that
coordinates the β- and γ-phosphates in GTP. Upon nucleotide hydrolysis the P-loop does
not significantly change conformation, but stays bound to the remaining phosphate
groups. β- and γ- subunits are usually anchored to the membrane by covalently attached
fatty acids. Gβγ can also directly participate in signal transduction. It activates a wide
variety of signaling proteins including several isoforms of Adenylate Cyclase.
Figure 2: Close view of G-protein with loops and subunits

Small G protein
These proteins belong to a large superfamily referred to as small G proteins based on
their low Mr of 20,000 to 35,000. The small G proteins, like the heterotrimeric G
proteins, bind guanine nucleotides, possess intrinsic GTPase activity and cycle through
GDP- and GTP-bound forms. One unifying feature of the various classes of G protein is
that the binding of GTP versus GDP dramatically alters the affinity of the protein for
some target molecule, apparently by inducing a large conformational change. Small G
proteins appear to function as molecular switches that control several cellular processes.
Examples of small G-protein: These are the following cellular actions which are
performed by small G-protein.
Table 1: Classification of small G-proteins on the basis of their functions
Sl. Class Proposed cellular function

no.
1. Ras Signal transduction (control of growth factor and
MAP- kinase pathway)
2. Rac, CDC42 Signal transduction (control of cellular stress
responses and MAP-kinase pathways)
3. Rab Localized to synaptic vesicle, where it regulates
vesicle trafficking and exocytosis
4. Rho Assembly of cytoskeleton structures (e.g. actin
microfilament)
5. ARF ADP-ribosylation
6. EFTU Associated with ribosomes where it regulates
protein synthesis
7. Ran Nuclear-cytoplasmic trafficking of RNA and
protein

Conformational changes occur in G protein during nucleotide exchange:
When G- protein couple receptor comes in contact of ligand then some conformational
changes occurs in GPCR; then the activated GPCR interact with G-protein, causes
conformational change in alpha subunit of G-protein. Due to conformational change, the
G-protein that bound to GDP prior to activation get exchange by GTP through which the
three switch regions on the α- subunit close to the nucleoside triphosphate, generating the
active conformation.
Figure 3: Conformational change in G-protein

Nature of G protein: G-Protein generally found in two states- active form and unactive
form.
In unactivated state, the guanylyl nucleotide bound to the G-Protein is GDP. In this form,
the G-protein exists as a heterotrimer consisting of α, β and γ subunits in which the α
subunit (Gα) binds to nucleotide. The role of the hormone- bound receptor is to catalyze
the exchange of GTP for GDP. Thus, inactive G-protein converted to active form and Gα
subunit has no much affinity for Gβγ subunits, hence α-subunit dissociated from βγ-
subunits. Now this is called activated state of G-protein. In activated state, Gα subunit
stimulate effector protein such as adenylyl cyclase that lead to production of second
messenger cAMP which may activate one or more signalling molecules. Other effectors
are cGMP phosphodiesterase, phospholipase C-β. After dissociation from the Gα subunit,
the βγ complex also has a signaling function and it can couple to at least four different
types of effectors: PLC-β, K+ ion channels, adenylyl cyclase, and PI 3-kinase.
Figure 4: Regulation of G- protein activation and inactivation

After certain amount of time passed , the Gα turn themselves off by hydrolysis of bound
GTP to GDP and inorganic phosphate (Pi). This results in conformational change caused
decrease in affinity for effector and increase in affinity for βγ complex, thus Gα subunit
dissociate from effector and associate with βγ subunit for the reformation of inactive
form of G-protein.
Figure 5: The mechanism of receptor-mediated activation (or inhibition) of effectors by means of heterotrimeric G proteins
Heterotrimeric G proteins come in five flavors, Gs, Gq, Gtα, Gi, and G12/13. This
classification is based on the Gα subunits and the effectors to which they couple. The
particular response produced by an activated GPCR depends on the type of G protein
with which it interacts, although some GPCRs can interact with different G proteins and
trigger more than one physiologic response.
1. Gs family members couple receptors to adenylyl cyclase. Adenylyl cyclase is
activated by GTP-bound Gs subunits.

2. Gqα, Gq family members contain Gα subunits that activate PLC-β. PLC-β hydrolyzes
phosphatidylinositol bisphosphate, producing inositol trisphosphate and
diacylglycerol.
3. Transducin (Gtα), a variant of Giα, which transduces visual stimuli by coupling the
light-induced conformational change of rhodopsin to the activation of a specific
phosphodiesterase, which then hydrolyzes cGMP to GMP. This cGMP-
phosphodiesterase (cGMP-PDE) is an αβγ2 heterotetramer that is activated by the
displacement of its inhibitory subunits (PDE) by their tighter binding to Gtα . GTP. A
cation-specific transmembrane channel that is held open by the binding of cGMP
closes on the resulting reduction in [cGMP], thereby triggering a nerve impulse
indicating that light has been detected.
4. Gi , Activated Gi subunits function by inhibiting adenylyl cyclase
5. Golf, a variant of Gsα, which is expressed only in olfactory sensory neurons and
participates in odorant signal transduction.
6. G12αand G13α, G12/13 members are less well characterized than the other G protein
families although their inappropriate activation has been associated with excessive cell
proliferation and malignant transformation.
Thus heterogeneity in G proteins occurs in the β and γ subunits as well as in the α

subunits. In fact, 21 different α subunits, 6 different β subunits, and 12 different γ
subunits have been identified in humans, some of which appear to be ubiquitously
expressed whereas others are expressed only in specific cells.Thus, a cell may contain
several closely related G proteins of a given type that interact with varying specificities
with receptors and effectors.This complex signalling system presumably permits cells to
respond in a graded manner to a variety of stimuli.
Role of G protein in signal transduction
Since, organs or cells in order to do their function properly within an organism must have
capacity to respond signals from distant cells as well as from its local environment. Thus
the process in which information carried by extracellular messenger molecules is
translated into changes that occur inside a cell is referred to as signal transduction. In
this process:

• Cells in different organs communicate with one another through extracellular
signalling molecules released by one set of cells and received by the other.
• Not all molecules can pass through the lipid bilayer of a Cell, and so signal
transduction systems are used in order to transmit an external signal to the cell interior.
G-proteins play key role in signal transduction with the help of G- protein couple receptor
which abbreviated as GPCR.
G- protein couple receptor: GPCR comprise a large protein family of transmembrane
receptors that sense molecules outside the cell and activate inside signal transduction
pathways and ultimately, cellular responses. G protein-coupled receptors are found only
in eukaryotes, including yeast, choanoflagellates and animals. The ligands that bind and
activate these receptors include light-sensitive compounds, odors, pheromones,
hormones, and neurotransmitters, and vary in size from small molecules to peptides to
large proteins. G protein-coupled receptors are involved in many diseases, and are also
the target of approximately 40% of all modern medicinal drugs.
Classification of GPCR: The exact size of the GPCR superfamily is unknown but nearly
800 different human genes (or ≈4% of the entire protein-coding genome) have been
predicted from genome sequence analysis. Although numerous classification schemes
have been proposed, the superfamily is classically divided into three main classes (A, B,
and C) with no detectable shared sequence homology between classes. The largest class
so far is class A, which accounts for nearly 85% of the GPCR genes. Of class A GPCRs,
over half of these are predicted to encode olfactory receptors while the remaining
receptors are liganded by known endogenous compounds or are classified as orphan
receptors. Despite the lack of sequence homology between classes, all GPCRs share a
common structure and mechanism of signal transduction.
In general, GPCRs can be classified into 5 classes based on sequence homology and
functional similarity:

G- Protein Coupled
Receptor
Figure 6: Classification of G-protein
Role of G- protein coupled receptor: GPCRs are involved in a wide variety of

physiological processes. Some examples of their physiological roles are as follows:
1. The visual sense: Rhodopsine which is complex of opsins and chromophore 11-
cis retinal, use a photoisomerization reaction to translate electromagnetic radiation
into cellular signals due to the conversion of 11-cis-retinal to all-trans-retinal.
2. The sense of smell: receptors of the olfactory epithelium bind odorants (olfactory
receptors) and pheromones (vomeronasal receptors)
3. Behavioral and mood regulation: receptors in the mammalian brain bind several
different neurotransmitters, including serotonin, dopamine, GABA, and glutamate
4. Regulation of immune system activity and inflammation: Chemokine receptors
bind ligands that mediate intercellular communication between cells of the
immune system; receptors such as histamine receptors bind inflammatory
mediators and engage target cell types in the inflammatory response
5. Autonomic nervous system transmission: Both the sympathetic and
parasympathetic nervous systems are regulated by GPCR pathways, responsible
for control of many automatic functions of the body such as blood pressure, heart
rate, and digestive processes
6. Cell density sensing: A novel GPCR role in regulating cell density sensing.
7. Homeostasis modulation (water balance).

Mechanism of action: When a ligand binds to the GPCR it causes a conformational
change in the GPCR, which allows it to act as a guanine nucleotide exchange factor
(GEF). The GPCR then activate an associated G-protein by exchanging its bound GDP
for a GTP. The G-protein's α subunit, together with the bound GTP then dissociate from
the β and γ subunits to further affect intracellular signaling proteins or target functional
proteins directly depending on the α subunit type (Gαs, Gαi/o, Gαq/11, Gα12/13).
There are two principal signal transduction pathways followed by the G protein-coupled
receptors:
• the cAMP signal pathway
• the Phosphatidylinositol signal pathway.
Adenylate Cyclase regulated by Gs and Gi type of Gα

The adenylate cyclase pathway regulated by both stimulatory and inhibitory subunits of
G protein Gs and Gi respectively.
In case of stimulatory subunit, Gα have intrinsic GTPase activity, which is used to
hydrolyze bound GTP to GDP and Pi. This hydrolysis reaction is slow, however,
requiring from second to minutes. Thus the GTP form of Gα is able to activate
downstream components of signal transduction pathway before GTP hydrolysis that
deactivates the subunit. In essence, the bound GTP acts as a built in clock that
spontaneously resets the Gα subunit after a short time period. After GTP hydrolysis and
release of Pi, the GDP –bound form of Gα then reassociates with Gβγ to re-form the
inactive heterotrimeric protein. Since Gα hydrolyzes its bound GTP at a characteristic
rate, it functions as a molecular clock that limits the length of time that both Gα. GTP
and Gβγ can interact with their effectors.
On the other hands, Gβγ can also directly participate in signal transduction by activation
of wide variety of signaling proteins including several isoforms of AC, certain Na+, K+
and Ca2+-specific ion channels, various protein tyrosine kinases and phospholipase C- β
(PLC-β; a component of the phospho-inositide signaling system; Section. Gβγ thereby
provides an important source of cross talk between signaling systems.
“Several types of ligand–GPCR complexes may activate the same G protein.This occurs,
for example, in liver cells in response to the binding of the corresponding hormones to

glucagon receptors and to β-adrenergic receptors. In such cases, the amount of cAMP
produced is the sum of that induced by the individual hormones.”
Figure 6: Mechanism of receptor-mediated activation/inhibition of Adenylate Cyclase.
In case of inhibitory type of α-subunit, some ligand–GPCR complexes inhibit rather than
activate AC. These include the α2-adrenergic receptor and receptors for somatostatin and
opioids. The inhibitory effect is mediated by “inhibitory” G protein, Gi, which may have
the same β and γ subunits as does “stimulatory” G protein, Gs, but has a different α
subunit,Giα (41 kD). Gi acts analogously to Gs in that on binding to its corresponding
ligand–GPCR complex, its Giα subunit exchanges bound GDP for GTP and dissociates
from G. However, Giαinhibits rather than activates AC, through direct interactions and
possibly because the liberated Gβγ binds to and sequesters Gsα. The latter mechanism is
supported by the observation that liver cell membranes contain far more Gi than Gs. The
activation of Gi in such cells would therefore release enough Giα to bind than available
Gs.

Cholera Toxin Stimulates Adenylate Cyclase by Permanently Activating Gsα:
The major symptom of cholera, an intestinal disorder caused by the bacterium Vibrio
cholerae, is massive diarrhea that, if untreated, frequently results in death from
dehydration. This dreaded disease is not an infection in the usual sense since the vibrio
neither invades nor damages tissues but merely colonizes the intestine, much like E. coli.
The catastrophic fluid loss that cholera induces (often over 6 liters per hour!) occurs in
response to a bacterial toxin. Indeed, merely replacing cholera victims’ lost water and
salts enables them to survive the few days necessary to immunologically eliminate the
bacterial infestation.
Cholera toxin (CT; also known as choleragen) is an 87-kD protein of subunit
composition AB5 in which the B subunits (103 residues each) form a pentagonal ring to
which the A subunit (240 residues) is bound its A subunit is cleaved at a single site by a
bacterial protease to yield two fragments, A1 (the N-terminal ~195 residues) and A2 (the
C-terminal ~45 residues), that remain joined by a disulfide bond. In the cytoplasm, A1
catalyzes the irreversible transfer of the ADP–ribose unit from NAD to a specific Arg
side chain of Gs. This reaction is greatly accelerated by the interaction of A1 with the
small Ras-like G protein ADP- ribosylation factor (ARF) in complex with GTP, which
normally functions to prime the formation of clathrin-coated vesicles.

Figure 7: Mechanism of action of cholera toxin. The cholera toxin’s A1 fragment in complex with ARF. GTP catalyzes the
ADP- ribosylation of a specific Arg residue on Gsα by NAD+, thereby rendering this subunit incapable of hydrolysing GTP.
ADP-ribosylated Gsα GTP can activate AC but is incapable of hydrolyzing its bound
GTP. As a consequence, the AC remains “locked” in its active state. The epithelial cells
of the small intestine normally secrete digestive fluid (an – HCO3- rich salt solution) in
response to small increases in [cAMP] that activate intestinal Na+ pumps through their
phosphorylation by PKA. The ~100-fold rise in intracellular [cAMP] induced by CT
causes these epithelial cells to pour out enormous quantities of digestive fluid, thereby
producing the symptoms of cholera. Gsα and Giα are members of a family of related
proteins, many of which have downstream effectors other than AC.
Termination of GPCR and interrupt binding with G protein
The hormone-bound activated receptor must be reset as well to prevent the continuous
activation of G proteins. This resetting is accomplished by two processes.
First, the hormone dissociates, returning the receptor to its initial, unactivated states. The
likelihood that the receptor remains in its unbound states depends on the concentration of
hormone.
Second, the hormone- receptor complex is deactivated by the phosphorylation of serine
and threonine residues in the carboxyl-terminal tail of the receptor kinase (also called G
protein receptor kinase 2, GRK 2) phosphorylates the carboxyl terminal tail of hormones-
receptor complex but not the unoccupied receptor. Finally, the molecule β-arrestin binds
to the phosphorylated receptor and further diminish its ability to activate G-protein.
Figure-8: Signal termination

Human Diseases Linked to the G Protein Pathway
There are so many human diseases are related to disfunction of G-protein and G-protein
coupled receptor which may be due to mutation in the genes that encode for these
proteins. Some of them are given in the following table:
Table 2: Disease due defective G-protein
Sl. Disease Defective G-protein

No.
1. Albright’s hereditary osteodystrophy and GSα
pseudohypathyroidism
2. McCune-Albright syndrome Gsα
3. Pituitary, thyroid tumors(gsp oncogene) Gsα
4. Adernocortical, ovarian tumors (gip oncogene) Giα
5. Combined precocious puberty and Gsα
pseudohypoparathyroidism
Table 3: Disease due to defective G-protein couple receptor

Sl. Disease Defective G- protein couple
No. receptor
1. Family hypocalciuric hypercalcemia Human analogue of BoPCAR1
receptor
2. Neonatal severe hyperparathyroidism Human analogue of BoPCAR1
(thyroid adenomas) receptor
3. hyperparathyroidism (thyroid Thyrotrophin receptor
adenomas)
4. Familial male precocious puberty Luteinizing Hormone receptor
5. X-linked nephrogenic diabetes V2 vasopressin recept
inspidus
6. Retinitis pigmentosa Rhodopsine receptor
7. Color blindness, spectral sensitivity Cone opsin receptor
variations
8. Familial glucocorticoid deficiency Adernocorticotrophic hormone
and isolated glucocorticoid (ACTH) receptor
deficiency
The mechanism for transmitting signals across the plasma membrane by G proteins is of
ancient evolutionary origin and is highly conserved.

G Proteins often Require Accessory Proteins to Function
The proper physiological functioning of a G protein often requires the participation of
several other types of proteins which are described below:
1. A GTPase-activating protein (GAP), which as its name implies, stimulates its
corresponding G protein to hydrolyze its bound GTP. This rate enhancement can be >
2000-fold. The downstream effectors of Gtα and Gqt, cGMP-PDE and PLC-β
respectively, exhibit GAP activities toward Gtα and Gqα (which otherwise would
hydrolyze GTP at physiologically insignificant rates), but AC does not exhibit GAP
activity toward either Gsα or Giα. However, in humans, a diverse family of 37 RGS
proteins (for regulators of G protein signaling) function as GAPs for Gα subunits by
binding most avidly to them when they are in the transition state conformation for
hydrolyzing GTP.
2. A guanine nucleotide exchange factor [GEF; alternatively guanine nucleotide releasing
factor (GRF)], which induces its corresponding G protein to release its bound GDP. The
G protein subsequently binds another guanine nucleotide (GTP or GDP, which most G
proteins bind with approximately equal affinities), but since cells maintain a GTP
concentration that is 10-fold higher than that of GDP, this, in effect, exchanges the bound
GDP for GTP. For heterotrimeric G proteins, the agonist–GPCR complexes function as
GEFs.
3. A guanine nucleotide-dissociation inhibitor (GDI). A Gβγ may be regarded as its
associated Gα’s GDI because GDP dissociates slowly from isolated Gα subunits but is
essentially irreversibly bound by heterotrimers.
Interesting facts:
1. G proteins are molecular switches that use GDP to control their signaling cycle. G
protein is inactive when GDP bounds. To activate the protein, the GDP is replaced
with GTP, and then G protein will deliver its signal.
2. The G protein system plays a central role in many signalling tasks, so it became
sensitive target for many drugs and toxins.

3. The diversity of GPCRs is observed not only by the multiplicity of stimuli to which
they respond, but also by the variety of intracellular signalling pathways they activate
Questions:
1. How G-protein activated when light fall on eye? What is the role of G-protein in
signalling in eye?
2. What are the effect of cholera toxin in cell signaling?
3. Which are the enzyme dependent on G-protein activation? How G-protein involve
in signal transduction?
References:
1. http://probis.cmm.ki.si/examples.php
2. Siegel GJ, Agranoff BW, Albers RW, et al., editors.Basic Neurochemistry:
Molecular, Cellular and Medical Aspects. 6th edition, chapter 20, table 20-2,
Philadelphia: Lippincott-Raven; 1999.
3. Berg, J., Tymoczko, J. and Stryer, L. Biochemistry. 5th edition, chapter-15
Cells
5. http://www.wiley.com/college/pratt/0471393878/student/animations/signal_transd
uction
6. Voet, D. and Voet, J. G. Biochemistry. 4th edition. Chapter-19: Signal
transduction
7. http://www.rcsb.org/pdb/101/motm.do?momID=58
8. http://jcs.biologists.org/content/116/24/4867

Cell Signalling Notes Merit Life Sciences1

Uploaded by

Copyright:

Available Formats

Cell Signalling Notes Merit Life Sciences1

Uploaded by

Document Information

Original Description:

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Cell Signalling Notes Merit Life Sciences1

Uploaded by

Copyright:

Available Formats

How does cell signaling work?

How does cell signaling work?

What are the main stages of cell signaling?

What are the main stages of cell signaling?

MERIT LIFE SCIENCES ONLINE Coaching for CSIR NET JRF LIFE SCEINCES

UNIT 4 MERIT LIFE SCIENCES

Cell signaling can be divided into 3 stages:

Extracellular fluid Cytoplasm

1. Reception 2. Transduction 3. Response

Figure 2: Cell signalling stages

Figure 3: Representation of Signal Transduction pathway

Figure 4: Common second messengers

Figure 5: Action of cAMP-dependent protein kinase

4. Signal termination by protein phosphatase: Signal termination is final step of signal

Figure 6: Signal amplification Pathways

Tissue Hormones Response

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

Receptor tyrosine kinases

Figure 1: Examples of receptors

Receptor types based on cellular location:

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

Estrogen Plasma membrane

Hsp 90 Receptor dimer

Inactive estrogen HAT

Figure 2: Estrogen receptor action

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

Structure of nuclear receptors: Nuclear receptors constitute a superfamily of dimeric

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

C-terminal domain (F): It is highly variable in sequence between various nuclear

Figure 3: Structural Organization of Nuclear Receptors (estrogen receptor)

Mechanism of action: The intracellular nuclear receptors respond to small hydrophobic

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

Figure 4: Model of hormone-dependent gene activation by a homodimeric nuclear receptor.

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

Figure 6 (a): Schematic diagram of G protein–coupled receptors

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

Epinephrine Binds to Several Different G Protein–Coupled Receptors. All epinephrine

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

1. Cyclic nucleotides (cAMP and cGMP)

We will discuss about all of them in the upcoming lectures.

Cyclic adenosine monophosphate (cAMP)

Figure 1: Cyclic Adenosine monophosphate or 3'-5'-cyclic adenosine monophosphate

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

Figure 2: Regulation of protein kinase A

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

Figure 3: Cyclic AMP inducible gene expression

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

7. Mitochondrial Biogenesis: cAMP/PKA has key role in mitochondrial compartment

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

Figure 5: Action of cAMP in our body

Mechanism of Regulation of cAMP: When stimulatory hormone binds to G-protein

INITITAIVE OF SCHOLARS & IIT BHU PROFESSOR

Synthesis and Degradation of cAMP: cAMP is a cyclic nucleotide which serves as an