CELL SIGNALLING

• Cell signaling is part of a complex system of communication that governs basic cellular
activities and coordinates cell actions.
• The function of communicating with the environment is achieved through a number of
pathways that receive and process signals which originates from the external
environment or from other cells within the organism or from different regions within
the cell.
• Cells deprived of appropriate survival signal will undergo apoptosis.
• The ability of cells to perceive and correctly respond to their microenvironment is the
basis of development, tissue repair, immunity as well as maintaining normal tissue
homeostasis.
• Errors in cellular information processing are responsible for diseases such as cancer,
autoimmune conditions and diabetes.

DEFINITIONS:

• Signaling: Cell-cell communication via signals.

• Signal transduction: Process of converting extracellular signals into intra-cellular
responses.
• Ligand: The signaling molecule.
• Receptors: Bind specific ligands and in turn activate one or more intracellular
pathways. These pathways depend on intracellular signaling proteins which process
the signal and transmit the signal to appropriate intracellular targets. The targets at the
end of signaling pathways are called effector proteins.
• Intercellular signaling: Communication between cells.
• Intracellular signaling: Communication within the cell, responding to extracellular and
intracellular stimuli.

INTRACELLULAR SIGNALING PATHWAY ACTIVATED BY AN EXTRACELLULAR SIGNAL

MOLECULE:

Cells receiving signals undergo 3 processes:

1. Reception
2. Transduction
3. Response

• Chemical signals are released by a signaling cell and received by target cell.
• Target cells have proteins called receptors which bind to signaling molecule and cause
response.
• The receptor activates one or more intracellular signaling pathways, involving a series
of signaling proteins.
• Finally, one or more of the signaling proteins alter the activity of effector proteins and
thereby the behavior of the cell.
Signaling molecules (Ligands) & receptors are specific to each other. A receptor will
typically bind only to its specific ligand and vice-versa.

FORMS OF SIGNALLING:

• Contact dependent signalling

• Paracrine signalling
• Endocrine signalling
• Autocrine signalling

CONTACT DEPENDENT SIGNALING:

• Also called as juxtacrine signaling / Direct activation.

• Contact dependent signaling requires cells to be in direct membrane - membrane
contact.
• This allows a group of cells to co-ordinate their response to a signal that only one of
the cell may have received.
• This is important during development and in immune responses.
PARACRINE SIGNALLING:

• Signals that act locally between cells that are close together are called paracrine
signals.
• Paracrine signals move by diffusion through the extracellular matrix.
• These types of signals usually elicit quick responses that last only a short amount of
time.
• In order to keep the response localized, paracrine ligands are usually quickly
degraded by enzymes or removed by neighboring cells.

• The transfer of signals between nerve cells is one example of paracrine signaling.
• The tiny space between nerve cells where signal transmission occurs is called a
synapse.
• Signals are propagated along nerve cells by fast-moving electrical impulses.
• When these impulses reach the end of one nerve cell, chemical ligands called
neurotransmitters are released into the synapse by the presynaptic cell.
• The neurotransmitters diffuse across the synapse.
• The small distance between nerve cells allows the signal to travel quickly, which
enables an immediate response.
• When the neurotransmitter binds to the receptor on the surface of the postsynaptic
cell, the next electrical impulse is launched.
• The neurotransmitters are degraded quickly or are reabsorbed by the presynaptic
cell so that the recipient nerve cell can recover quickly and be prepared to respond
rapidly to the next synaptic signal.
ENDOCRINE SIGNALING:

• Signals from distant cells are called endocrine signals, and they originate from
endocrine cells. The ligands released in endocrine signaling are called hormones.
• In the body, many endocrine cells are located in endocrine glands, such as thyroid
gland, hypothalamus and pituitary gland.
• Hormones travel the vast distances between endocrine cells and their target cells via
the bloodstream, which is a relatively slow way to move throughout the body.
• Thus these types of signals produce a slower response but have a long-lasting effect.
• Only those cells or tissues that contain the appropriate receptor for the hormone
elicit a hormonal response.

AUTOCRINE SIGNALING:

• When a cell responds to its own signaling molecule, it is called autocrine signaling
• This means the signaling cell and the target cell are one and the same.
• This type of signaling often occurs during the early development of an organism to
ensure that cells develop into the correct tissues and take on the proper function.
• Autocrine signaling also regulates pain sensation and inflammatory responses.
• If a cell is infected with a virus, the cell can signal itself to undergo programmed cell
death, killing the virus in the process.
TYPES OF RECEPTORS:

• Extracellular signal molecules bind to specific receptors.

• Receptors are protein molecules in the target cell or on its surface that bind to
ligands.

There are two types of receptors;

1. Internal receptors / intracellular / cytoplasmic receptors

2. Cell-surface receptors / transmembrane receptors:

• Enzyme-linked receptors
• Ion channel-linked receptors
• G-protein-linked receptors

INTRACELLULAR RECEPTORS:

• These are found in the cytoplasm/nucleus of target cells and respond to

hydrophobic ligand molecules that are able to travel across the plasma membrane.
• The signal molecule has to be small to diffuse across the plasma membrane and bind
to receptor proteins inside the target cell - either in the cytosol or nucleus.
• Once inside the cell, these molecules bind to proteins that act as regulators of mRNA
synthesis (transcription) to mediate gene expression.
• Gene expression is the cellular process of transforming the information in a cell’s
DNA into a sequence of amino acids, which ultimately forms a protein.
• When the ligand binds to the internal receptor, a conformational change (shape
change) is triggered that exposes a DNA-binding site on the receptor protein.
• The ligand-receptor complex moves into the nucleus, then binds to specific
regulatory regions of the chromosomal DNA and promotes the initiation of
transcription.
• Transcription is the process of copying the information in a cell’s DNA into a special
form of RNA called messenger RNA (mRNA); the cell uses information in the mRNA
to link specific amino acids in the correct order, producing a protein.
• Thus, when a ligand binds to an internal receptor, it can directly influence gene
expression in the target cell.
 Hydrophobic signaling molecules diffuse across the plasma membrane and
interact with intracellular receptors in the cytoplasm. Many intracellular receptors
are transcription factors that interact with DNA in the nucleus and regulate gene
expression.

CELL-SURFACE RECEPTORS:

• These are integral proteins that bind to external signaling molecules.

• The receptors are found around the plasma membrane and perform signal
transduction, in which an extracellular signal is converted into an intercellular signal
that alter the behaviour of cell.
• Errors in the protein structures of certain receptor molecules have been shown to
play a role in hypertension (high blood pressure), asthma, heart disease and cancer.
 Hydrophilic signaling molecules typically work by binding to the extracellular portion of a
receptor protein. The signal is then transduced across the membrane.

Each cell-surface receptor has three main components:

• an external ligand-binding domain, or extracellular domain;

• a hydrophobic membrane-spanning region; and
• an intracellular domain.

Cell-surface receptors are involved in most of the signaling in multicellular organisms.

There are three general categories of cell-surface receptors:

• Enzyme-linked receptors
• Ion channel-linked receptors
• G-protein-linked receptors

Enzyme-linked receptors:

• Enzyme-linked receptors normally have large extracellular and intracellular domains,

that are associated with an enzyme
• When a ligand binds to the extracellular domain of an enzyme-linked receptor, a signal
is transferred through the membrane, activating the enzyme.
• Activation of the enzyme sets off a chain of events within the cell that eventually leads
to a response.
• One example of an enzyme-linked receptor is the tyrosine kinase receptor.
• A kinase is an enzyme that transfers phosphate groups from ATP to another protein.
• The tyrosine kinase receptor transfers phosphate groups to tyrosine molecules.
• First, signaling molecules binds to the extracellular domain of two nearby tyrosine
kinase receptors.
• The two neighboring receptors then bond together, or dimerize.
• Phosphates are then added to tyrosine residues on the intracellular domain of the
receptors (phosphorylation).
• The phosphorylated residues can then transmit the signal to the next messenger
within the cytoplasm.
Ion channel-linked receptors:

• Also called transmitter gated ion channels or ionotropic receptors.

• Ion channel-linked receptors bind to a ligand and open a channel through the
membrane that allows specific ions to pass through.
• When a ligand binds to the extracellular region of the channel, there is a
conformational change in the protein’s structure that allows ions such as sodium,
calcium, magnesium, or hydrogen to pass through.
• An example of an ion channel-linked receptor is found in neurons. When
neurotransmitters bind to these receptors, a conformational change allows sodium
ions to flow across the cell membrane, causing a change in the membrane potential.
G-protein coupled receptors:

• G-protein-linked receptors bind a ligand and activate a membrane protein called a G-

protein, which then interacts with either an ion channel or an enzyme in the
membrane.
• The G protein-coupled receptors are characterized by seven membrane-spanning α
helices
• Approximately about 45% of all pharmaceutical drugs are known to target GPCRs
• Mammalian G protein complexes are made up of - 20 alpha (α); 6 beta (β); 12
gamma (γ) subunits.
• Cell signaling using G-protein-linked receptors occurs as a cycle. Once the ligand
binds to the receptor, the resultant shape change activates the G-protein, which
exchanges GDP for GTP (GDP molecule associated with the α subunit).
• The subunits of the G-protein then split into α and βγ subunits. One or both of these
G-protein fragments may be able to activate other proteins in the cell.
• After a while, the GTP on the active α subunit of the G-protein is hydrolyzed to GDP
and the βγ subunit is deactivated. The subunits re-associate to form the inactive G-
protein and the cycle begins again.
• G-protein linked receptors are used in many physiological processes including those
for vision transduction, taste, and regulation of immune system and inflammation.
TRANSDUCTION:

• The transduction stage of signaling is usually a multi-step pathway and these

pathways amplify the signal and provide coordination and regulation.
• A small number of signal molecules can produce a large cellular response.

Protein Phosphorylation and Dephosphorylation:

• In many pathways, the signal is transmitted by a cascade of protein phosphorylation.

Protein kinases transfer phosphates from ATP to protein, a process called

phosphorylation.

Protein phosphatases remove the phosphates from proteins, a process called

dephosphorylation.

• This phosphorylation and dephosphorylation system acts as a molecular switch,

turning activities on and off or up or down, as required.
Messengers:

• The extracellular signal molecule (ligand) that binds to the receptor is a pathway's
"first messenger".
• Second messengers are small, non-protein, water-soluble molecules or ions that
spread throughout a cell by diffusion.
• Second messengers participate in pathways initiated by GPCRs and Receptor tyrosine
kinase (RTK)

Second messengers:

• May be formed and inactivated by enzymatic reactions

• May be released from storage sites
• Are cytosolic or membrane-localized
• Activate signaling enzymes
• Allow signal amplification
• Are produced and become active in a timely and locally controlled way.

Types of Signalling Ligands:

A. Ligands that bind to cell-surface receptors:

1. Neurotransmitters (NT), i.e. norepinephrine, histamine

2. Peptide hormones (P), i.e. insulin
3. Growth factors (GF),
4. Lipophilic signaling molecules, i.e. prostaglandins

B. Ligands that bind to intracellular receptors:

Lipid soluble hormones that diffuse across the plasma membrane and interact with
receptors in the cytosol or nucleus. i.e. steroids, thyroxine, retinoic acid, nitric oxide.

Types of Signalling ligands: (simpler classification)

1. Small Hydrophobic Ligands

2. Water-Soluble Ligands
3. Other Ligands

Small hydrophobic ligands:

Small hydrophobic ligands can directly diffuse through the plasma membrane and
interact with internal receptors. Important members of this class of ligands are the
steroid hormones.
Water-soluble ligands:

• Water-soluble ligands are polar and, therefore, cannot pass through the plasma
membrane unaided. Instead, most water-soluble ligands bind to the extracellular
receptors.
• These water soluble ligands are quite diverse and include small molecules, peptides,
and proteins

Other Ligands:

• Nitric oxide (NO) is a gas that also acts as a ligand.

• It is able to diffuse directly across the plasma membrane; one of its roles is to
interact with receptors in smooth muscle and induce relaxation of the tissue.
• NO has a very short half-life; therefore, it only functions over short distances.

SECOND MESSENGERS:

There are 3 major classes of second messengers:

1. Cyclic nucleotides (e.g., cAMP and cGMP)

2. Inositol trisphosphate (IP3) & diacylglycerol (DAG)
3. Calcium ions (Ca2+)

CYCLIC NUCLEOTIDES:

Cyclic AMP (cAMP)

Some of the hormones that achieve their effects through cAMP as a second messenger –
adrenaline, glucagon, luteinizing hormone (LH), FSH, MSH, PTH

• Cyclic AMP is synthesized from ATP by the action of the enzyme adenylyl cyclase.
• Binding of the hormone (Ligand) 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):

1. Changing the molecular activities in the cytosol, often using Protein Kinase A (PKA)
2. Turning on a new pattern of gene transcription.

Cyclic GMP (cGMP)

• Cyclic GMP is synthesized from the nucleotide GTP using the enzyme guanylyl
cyclase.
• Cyclic GMP serves as the second messenger for - atrial natriuretic peptide (ANP),
nitric oxide (NO)
• 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.

INOSITOL TRISPHOSPHATE & DIACYLGLYCEROL:

• Peptide and protein hormones like vasopressin, TSH, neurotransmitters like GABA
bind to G protein-coupled receptors (GPCRs) that activate the intracellular enzyme
phospholipase C (PLC).
• PLC hydrolyzes phospholipids - specifically phosphatidylinositol 4,5-bisphosphate
(PIP2) which is found in the inner layer of the plasma membrane.

Hydrolysis of PIP2 yields two products:

1. Diacylglycerol (DAG) acts via Protein Kinase C (PKC) - a calcium-dependent kinase

2. Inositol-1,4,5-trisphosphate (IP3)

• DAG remains in the inner layer of the plasma membrane.

• It recruits Protein Kinase C (PKC) - a calcium-dependent kinase that phosphorylates
many other proteins that bring about the changes in the cell.
• IP3 - diffuses through the cytosol and binds to receptors on the endoplasmic reticulum
causing the release of calcium ions (Ca2+) into the cytosol.
• The rise in intracellular calcium triggers the response.

CALCIUM IONS:

• Calcium ions are the most widely used intracellular messengers.

• 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 - release of neurotransmitters at synapses,

activation of T cells and B cells - when they bind antigen with their antigen
receptors, apoptosis.

IP3 binds with receptors on SER (smooth endoplasmic reticulum) to stimulate the release
of calcium ions as a part of the amplification of a hormone's regulation of cellular
enzymes.

Termination of Signal:

• Signal response is terminated quickly by reversal of ligand binding.

• Inactivation mechanisms are an essential aspect of cell signaling.
• Unbound receptors revert to an inactive state.