SIGNALS & SIGNAL

TRANSDUCTION

dr. Tiwuk Susantiningsih, M.Biomed

Blok MBS1, FK UNILA
2014
INTRODUCTION.
Complex organism

Have special function.

Each cell contribute in an integrated way as the
body grows, differentiates, and adapts to
changing conditions, such as : hypoxia, in a hurry

Such integration requires communication that

carried out by
chemical messennger traveling from one cell
to others or
by direct contact with extracellular matrix or
with each cell
The goal of the signal is

to change actions carried out in target cells by

intracellular protein such as
metabolic enzymes
gene regulatory proteins
ion channels
cytoskeletal proteins.

Example:
Cells in condition of Hypoxia change their metabolic
to generate ATP from aerobic to be an aerobic
glycolysis.
In hypoxic condition cells express in a higher
concentration of a transcription factor protein
called HIF-1 (hypoxia inducible factor)

that regulate the expression of others

functional protein such as glycolytic enzymes,
VEGF (vascular endothelial growth factor, EPO
(erythropoetin).

all fuctional proteins act synergically to adapt

hypoxic condition to survive or to maintain
homeostatic millieu.
 Cell signaling by chemical
messengers
Chemical messenger
Also called signaling molecules
Transmit message between cells
They are secreted from one cell in response to a
specific stimulus
Travel to a target cell where they bind to a
specific receptor and elicit a response
In the nervous system, these chemical
messenger are called neurotransmitter
In the endocrine they are hormones
In the immune system they are called cytokines
Additional chemical-messenger (ch-m) are
retinoids, eicosanoids, and growth factors
According to the distance between the
secreting and target cell, ch-m are
classified as
endocrine : travel in the blood
paracrine : travel between nearby
cells
autocrine : act on the same cell or
on nearby cells of the
same type
Receptor and Signal Transduction

Receptor (radar, parabola) are protein

containing a binding site specific for a single
chemical messenger and another binding site
involved in transmitting the message (multiple
bindiing site)

The second binding site may interact with

another protein or with DNA (for steroid
hormone).

Type:
Plasma membrane receptors
Intracellular binding protein
Ionotrophic Receptor
 Plasma membrane receptor.
Span the plasma membrane and contain an
extracellular binding domain for the messenger.

Intracellular binding protein (intracellular

receptors) are for messenger able to diffuse into
the cell.

Most plasma membrane receptors are devided

into categories
Ion channel receptors
Tyrosine kinase receptors
Tyrosine-kinase associated receptors (JAK-
STAT )
Serine-threonin kinase receptors or
heptahelical receptors (proteins with seven
helices spanning the membrane)
When chemical messenger binds to receptor, the
signal it is carrying (the messages) must be
converted into an intracellular response.

The conversion is called signal transduction.

Signal transduction for intracellular receptors.

Most intracellular receptors are gene-specific
transcription factors (a protein that can bind to
DNA and regulate the transcription of certain
gene).
Signal transduction for plasma
membrane receptors.
Mechanism of signal transduction that follow the
binding of signaling molecules to plasma
membrane receptors include

Phosphorilation of receptors at tyrosin residues

(receptor tyrosine kinase activity)

Conformational changes in signal transducer

proteins (e.g. protein with SH domain, the
monomeric G protein Ras, heterotrimeric G
protein).Or increases the level of intracellular
second messengers.
Second messenger are non protein molecules
generated inside the cell in response to hormone
binding that continue to transmission of the
message.
Examples: 3,5-cyclic AMP (cAMP),
inositol triphosphate (IP3) and
diacylglycerol (DAG).

Signaling often requires a rapid response and rapid

termination of the message, which maybe achieved
by
Degradation of the messenger or second
messenger.
The automatic G protein clock (activated and
inactivated).
Deactivation of signal transduction kinases by
phosphatase or other means.
General Feature of Chemical Messenger
Signaling sequence:
The chemical messenger is secreted from a
specific cell in response to stimulus
The chemical messenger diffuses or is
transported through blood or other cellular fluid
to target cell
A receptor in the target cell specifically binds the
messenger
Binding of the messenger to the receptor elicit a
respons
The signal cease and is terminated

Chemical messenger elicit their response in the

target cell without being metabolized by the cell
The specificity of the response is dictated by
the type of receptor and its location.
Generally each receptor bind only one specific
chemical messenger.
Each receptor initiates a characteristic signal
transduction pathway that will ultimately activate
or inhibit certain processes in the cell.
Only certain cells (target cells) carry receptors for
that messenger and are capable of responding to
its message.

The means of signal termination is

an exceedingly important aspect of cell signalling,
failure to terminate a message contributes to a
number of diseases, such as cancer.
Chemical messenger system applied to the
Nicotinic Acetylcholine Receptor
Cell signaling, ilustrated with acetylcholine ,a
neurotransmitter that acts on nicotinic acetylcholine
receptors on plasma membrane of certain muscel
cells.

This system exhibits the classic features of chemical

messenger release and specificity of response.

Neurotransmitter are secreted from neurons in response to

an electrical stimulus called the action potential (a voltage
difference across the plasma membrane caused by changes
in Na+ and K+ gradients, that is propagated along a nerve)

The neurotransmitters diffuses across a synapse to another

excitable cell, where they elicit response
Acetylcholine (AC) = neurotransmitter at neuromuscular
junction, where it transmits a signal from a motor
nerves to a muscel fiber that elicits contraction of
fiber.

Before release AC is sequestered in vesicles clustered

near an active zone in the presynaptic membrane.
This membrane also has voltage-gated Ca++ channels
that open when the action potential reachs them
influx of Ca++.
Ca++ triggers fusion of vesicles with the plasma
membrane AC released into the synaptic cleft.
AC diffuses across the synaptic cleft to bind to plasma
membrane receptors on the muscle cell called
nicotinic acetylcholine receptors.
The receptor subunits are assembled around a
channel, which has funnel-shaped opening in the
center.
As AC binds to the receptor, a conformational change
opens the narrow portion of the channel/ the gate,
allowing Na+ to diffuse in and K+ to diffuse out.
(A uniform property of all receptors is that signal
transduction begins with conformational changes in
the receptor.)
The change in ion concentration activates a sequence
of events that eventually triggers the cellular
response contraction of the fiber.
Once AC secretion stops, the message is rapidly
terminated by acetylcholine esterase, an enzyme
located on the post synaptic membrane that cleaves
AC.
It is also terminated by diffusion of AC away from the
synaps
Rapid termination of message is a characteristic of
systems requiring a rapid response from target cell.

Endocrine, Paracrine and Autocrine.

The action of chemical messenger are often classified

as endocrine, paracrine and autocrine.
Each hormone is secreted by specific cell type
(generally in an endocrine gland), enters the blood
and exert its actions on specific target cell, which
may be some distance away.
In contrast to the endocrine hormones, paracrine
actions are those performed on nearby cells, and the
location of the cells plays a role in specificity of the
response.
Synaptic transmission by AC and other
neurotransmitters (sametimes called neurocrine
signaling) is an example of paracrine signaling.

Autocrine action involve a messenger acting on:

The cell from which it is secreted, or
The nearby cells that are the same type as the
secreting cells

Type of Chemical Messengers.

1. The nervous system
2. The endocrine system
3. The immune system
4. The eicosanoids
5. Growth factors
1. The Nervous system.

The nervous system secretes 2 types of messengers

Small molecule neurotransmitters/ biogenic
amines
Neuropeptides.

Neurotransmitter are nitrogen-containing molecules,

which can be amino acids or are derivatives of
amino acids (AC, -aminobutyrate).

Neuropeptides are usually small peptides (4 35 AA),

secreted by neurons, that act as neurotransmitters
at synaptic junctions or are secreted into the blood
to act as neurohormones.
2. The endocrine system

Hormones are defined as compounds, secreted from

specific endocrine cells, in endocrine glands, that
reach their target cells by transport through the
blood.
Example: insulin
Hormone secreted from the -cell of pancreas.

Classic hormones are devided in to the structural

categories of
Polypeptide hormones (insulin, glukagon)
Catecholamine (epinephrine = neurotransmitter)
Steroid hormones (derived from cholesterol)
Thyroid hormone (derived from tyrosine)
Many of these endocrine hormones also exert
paracrine and autocrine actions.

Some compounds normally considered hormones are

more difficult to categories, for examples:
Retinoids, which are derivatives of vitamin A
(retinol)
Vitamin D (derived from cholesterol) are usually
classified as hormones although they are not
synthesized in the endocrine cells.

3. The immune system

The messenger of the immune system called cytokines

, are small proteins with a MW approximately 20.000
daltons
Cytokines regulate a network of responses designed to
kill invading microorganisms.

The classes of cytokines:

Interleukins
Tumor necrosis factors
Interferons
Colony-stimulating factors

Cytokines are secreted by cells of the immune system

and usually alter the behavior of other cells in the
immune system by activating the transcription of
genes for proteins involved in the immune response.
4. The eicosanoids.

The eicosanoids are (paracrine and autocrine

functions):
Prostaglandins (PG)
Thromboxanes
Leukotrienes

They control cellular function in respon to injury.

The compounds are all derived from arachidonic acid, a
20-C polyansaturated fatty acid (FA) that is usually
present in cells as a part of the membrane lipid
phosphatidylcholine.
Although almost every cell in the body produces an
eicosanoid in response to injury, different cell
produce different eicosanoids.
Example: Vascular endothelial cell secrete the
prostaglandin PGI2 (prostacyclin) which act on
nearby smooth muscel to cause vasodilatation.

5. Growth factors

They are polypeptides that function through stimulation

of cellular proliferation.
Example: platelets aggregating at the site of injury to a
blood vessel secrete PDGF (platelet-derived growth
factor).
PDGF stimulates the proliferation or nearby smooth
muscle cells, which eventually form a plaque
covering the injury site.
Some growth factors are considered hormones, and
some have been called cytokines.
Each of the hundred of chemical messengers has its
own specific receptor, which will usually bind NO
OTHER MESSENGER.

I. INTRACELLULAR TRANSCRIPTION FACTOR

RECEPTORS.

Intracellular vs Plasma Membrane Receptors:

Intrcellular receptors can bind hydrophobic molecules

that able to diffuse through the palma membrane
into cells.
Extracellular receptors/ membrane receptors can
bind polar molecules such as peptide hormones,
cytokines, cathecolamines that cannot rapidly cross
the membrane plasma.
Most of the intracellular receptors for lipophylic
messenger are gene-specific transcription
factors.

A transcription factor is a protein that binds to

a specific site on DNA and regulates the rate
of transcription of a gene (synthesis of the
mRNA)

External signaling molecules binds to

transcription factors that bind to a specific
sequence on DNA and regulate the
expression of only certain gene;they are
called gene-specific or site specific
transcription factors
B. The Steroid hormone/Thyroid hormone
Superfamily receptors

Lipophilic hormons that use intracellular gene-specific

transcription factors include
Steroid hormone
Thyroid hormone
Retinoic acid
Vitamin D

Because these compounds are water insoluble they are

transported in the blood to serum albumin,
SHBG/steroid hormone binding globulin and TBG/
Thyroid hormone binding globulin.
The intracellular receptors for these hormones are
structurally similar

Primarily intracellular receptor found in the nucleus,

although some are found in the cytoplasm.

For example: Glucocorticoid receptor exist as

cytoplasmic multimeric complex ascociated with heat
shock protein

When the hormone cortisol binds, the receptor

undergoes a conformational change and dissosiates
from the heat shock protein, exposing a nuclear
translocation signal.
The receptor dimerize, and the complex (including
bound hormone) translocates to the nucleus, where
it binds to a pertion of the DNA called Hormone
Receptor Element/ HRE.

Example: glucocorticoid receptor binds to the

glucocorticoid response element/ GRE.

Most of the intracellular receptor reside principally in

the nucleus, and some of these are constitutively
bound, as dimers, to the response element in DNA
(e.g. thyroid hormone receptor).

Binding of the hormone changes its activity and its

ability to associate with or disassociate from DNA
II. Pasma Membrane Receptors and
Signal Transduction.

All plasma membrane receptor are proteins with

certain features in common
An extracellular domain that binds the chemical
messenger
One or more membrane spanning domains that are
-helix
An intracellular domain that initiates signal
transduction.

As the ligand binds to the extracellular domain of its

receptor, it causes a conformational change that is
communicated to the intracellular domain through
tha rigid -helix of the transmembrane domain.
The pathway of signal transduction for plasma
membrane have 2 major types of effect on cell
Rapid and immediate effect on cellular ion
level or activation and inhibition of enzymes
aktivasi/inhibisi enzim
Slower changes in the rate og gene expression
for a specific set of proteins
Often, a signal transduction pathway will diverge to
produce both kinds of effects.

A. Major Classes of Plasma Membrane Receptor

1. Ion channel receptors
2. Receptors that are kinases or bind kinases
3. Heptahelical receptors
1. Ion Channel Receptors.

The structure is similar to the nicotinic acetylcholine

receptor.
Signal transduction consist of the conformational
change when ligand binds
Most small molecule neurotransmitters and some
neuropeptide use ion channel receptors

2. Receptors that are kinases or bind kinases.

Their common feature is that the intracellular domain

of the receptor (or an associated protein) is a kinase
that is activated when the messenger binds to the
extracellular domain.
The receptor kinase phosphorilates an amino acid
residu on the receptor (autophosphorilation) or an
associated protein.
The message is propagated through signal transducer
proteins that bind to the activated messenger-
receptor complex (Grb2, STAT, Smad)

3. Heptahelical receptors

Heptahelical receptor contain 7-membrane spanning -

helices
They work through second messengers, which are
small non protein compouns such as c-AMP
Ligand of the receptor are
protein hormones
cytokines
neurotransmitter
Second messenger are present in low concentrations
so that their concentration, and hence the message
can be rapidly initiate and terminated.
B. Signal Transduction Through Tyrosine
Kinase Receptors

The receptors exist in membrane as monomer

with a single membrane spanning helix.

One molecule og growth factor generally binds

2 molecule receptors and promote dimer.

Once receptor dimer has formed the

intracellular tyrosine kinase domain of
receptor phophorylated each other
(autophosphorylation)

Phosphotyrosine risidues form specific binding

sitefor signal transducer protein
1 RAS and The MAP Kinase Pathway.

One of the domain of receptor contain a

phosphotyrosine residues form a binding site
for intracellular protein with specific 3-
dimensional structure =SH domain (Src
homology 2)
Src = Rous sarcoma virus
The adaptor protein Grb2 bound to a membran
phosphoinositide(contain SH domain)
The receptor bind growth factor
Ligand-receptor complex cause conformational
change in Grb2 that activate another binding
site=SH3 domain
Activated SH3 can bind protein=SOS, SOS is a guanine
nucleotide exchnge factor (GEF) for Ras, a
monomeric G protein located in the membrane
plasma

SOS activate exchange of GTP for GDP on Ras, causing

a conformational change in Ras that promote binding
to the protein Raf.

Raf is a serine protein kinase that also called MAPKKK

(mitogen activated protein kinase kinase kinase)

Raf begin a sequence of successive phosphorilation

step called phosphorylation cascade
The MAP kinase cascade terminates at gene
transcription factor, there by regulating transcription
or crtain genes involved in cell survival and
proliferation.
Many tyrosin kinase receptors (as well as hepta helical
recp) also have additional signaling pathway
involving phosphatidylinositol phosphates.

2. Phosphayidylinositol phosphat (PIP) in signal

transduction.

PIP serve 2 different functions

Fosfatidilinositol 45bisphosphates(PI-4,5-bisP)
devided into 2 products (second messengers).:
DAG/diacyl glycerol and inositol triphosphate/IP3
Phosphatidyllinositol 345 triphosphates (PI-3,4,5-
trisP) act as docking site on plasma membran for
signal transducer proteins
Fosfatidil inositol on plasma membrane Changed to be
PI-4,5-bisP by kinase enzyme.

PI-4,5-bisP catalyzed byphospholipase C isozyme

produce DAG dan IP3

Phospholipase C (PLC) activated by tyrosin kinase-

growth factor receptor.
Phospholipase C Isozyme activated by signal
transduction pathway of heptahelices- G protein
receptor

PI-4,5-bisP can be phosphorylated on their inositol

residue by phosphatidyl inositol 3 kinase produce PI-
3,4,5-trisP.

Both kind of PI act as docking site for protein that

contain domain pleckstrin homology (PH).
PI3 kinase contain SH2 domain that activated by
binding with phosphotyrosine site at tyrosin kinase
receptor.

3. Insulin receptor.

This receptors are member of tyrosin kinase receptor

Family. An example of diverge signal transduction.

Insulin receptor exist as dimer ( dan ).

Subunit phosphorylated each other when insulin bind
to the receptor activated the receptor.

Activated and phosphorylated receptor can binds dapat

protein IRS (insulin receptor substrate).
Then IRS protein phosphorilated on many site that can
bind proteins with SH2 domain.

One of the site bind Grb2 that activate Ras and MAP
kinase pathway.

Grb2 bind to PI-3,4,5-trisP in plasma membrane by

PH(pleckstrin homology) domain

In other site PI3 kinase will be activated

The third site can bind PLC and activated.

The signal pathway initiated by insulin-receptor
complex involving PI 3-kinase leads to activation of
kinase B.
PI3-kinase bind and phosphorylates PI-4,5-bisP in the
membrane to form PI-3,4,5-trisP.
PKB and PDK1 (phosphoinositide-dependent kinase-1)
are recruited to the membrane by theur PH domain,
wher PDK1 phosphorylates and activates kinaseB.

C. Signal transduction by JAK-STAT

receptors
Cytokines use this receptor to activate immune
system
Has no intrinsic kinase activity but bind to tyrosine
kinase JAK (Janus kinase)
Their signal transducer protein = STAT (signal
transducer and activator of transcription) , a
signal transducer and activator of
transcription.

D. Reseptor serin/treonin kinase

Transformated the growth factor receptor

superfamily and associated with Smad famili
protein .
Consist of transforming growth factor (TGF-
) , a cytokin for tissue repair, immune
regulation and cell proliferation, bone
morphogenetic proteins (BMPs) that control
proliferation.
E. Signal transduction through Heptahekical Receptor
Consist of:
1. Protein G heterotrimerik
2. Adenilat siklase , cAMP dan fosfodiesterase
3. Signal fosfatidil inositol by reseptor heptaheliks

1. Protein G
Binding of the hormon to receptor activete G protein
and then activates adenylate cyclase.
Hormones that can bind to are e.g. glucagon or
epinephrin.
When hormone binds the receptor leads to a
conformational change that cause GDP release and
GTP binding that cause subunit released
Alpha subunit will binds to enzyme target in the
membran= adenilat cyklase.
Adenilat cyklase will catalyze reactio that convert ATP
to c-AMP.

2. Adenilat siklase ,c-AMP, fosfodiesterase.

c-AMP will exert an effect inside the cell, such as
metabolic enzymes
Kinase A protein coul enter the nucleus and
phosphorylates gene-specific transcription CREB
(cAMP response element binding protein).

3. PI signaling by heptahelical receptors

Receptors bind to q isoform of protein G Subunit ,

and activate phospholipase C.
Phospholipase C hydrolyze PI-4,5-bisP to become 2
kind of second messengers DAG and IP3.
IP3 can attach/bind to sarcoplasmik reticulum and
endoplasmic reticulum and stimulate the release of
Ca2+
Ca2+ activates enzym that contain calcium-calmodulin
activate protein kinase.

DAG, still attach in membrane and activate protein

kinase C that trigger response: Phosphorilation of
proteins.

F. Changes in Response to Signals.

The Respon will be various.

Depend on the amount of the receptors
Down regulation
Degradation of the reseptor.
IV. Signal Termination.
Nerve signal should be terminated rapidly after the
hormone not produced anymore.
Stimulation of proliferation will be terminated slowly.
Signal for regilating diferentiation may persist through
out oer life time.
Some chronic desiases are caused by failure to
terminate a response at the appropriate time.
The first is the messengers no longer be secreted and
existing messengers will be catabolized.
Insulin taken up in to the liver and degraded.
cAMP catabolyzed by phosphodiesterase.
G protein hydrolyze GTP.
For kinase will be dephosphorylated by phosphatase
enzymes.