CELL SIGNALLING

Dr Saira Saad

1st YEAR
 CELL SIGNALLING

The survival of multi-cellular organisms depends on their ability to adapt

to a constantly changing environment. Intercellular communication
mechanisms are necessary requirements for this adaptation. The nervous
system and the endocrine system provide this intercellular, organism-wide
communication. There is a remarkable convergence of these two regulatory
systems. For example, neural regulation of the endocrine system is important
in the production and secretion of some hormones; many neurotransmitters
resemble hormones in their synthesis, transport, and mechanism of action; and
many hormones are synthesized in the nervous system.
Cell signalling can be classified into three distinct types based on the
distance over which the signalling molecule acts.
 In endocrine signalling, the signalling molecule (e.g. insulin) acts on
target cells distant from its site of synthesis in cells of an endocrine
organ (e.g. the pancreas). The endocrine cells secrete the signalling
molecule into the bloodstream (if an animal) or the sap (if a plant)
which carries it to the target cells elsewhere in the organism.
 In paracrine signaling, the signalling molecule affects only target
cells close to the cell from which it was secreted. The
communication from one nerve cell to another by chemical
neurotransmitters is an example of paracrine signaling.
 The third type of cell signaling is autocrine signaling, where a cell
responds to a molecule that it has produced itself.

Dr Saira Saad
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THE SIGNALLING MOLECULES:

The signaling molecules, ligands or hormones can be classified based on their
solubility and the location of their receptor
Classification
 Lipophilic hormones with intracellular receptors:
Small lipophilic (lipid-soluble) hormones diffuse across the plasma
membrane and then interact with intracellular receptors in the cytosol or
nucleus. The resulting hormone– receptor complex often binds to regions
of the DNA and affects the transcription of certain genes.
Examples:
 Steroid hormones e.g, the female sex hormones estrogen and
progesterone)
 Thyroxine
 Retinoic acid which is derived from vitamin A
 Vitamin D
 Lipophilic hormones with cell-surface receptors:
The principal lipophilic (lipid-soluble) hormones that bind to
receptors located in the plasma membrane are the prostaglandins.
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 Hydrophilic hormones with cell-surface receptors:

All hydrophilic (water-soluble) molecules (which cannot diffuse
across the hydrophobic interior of the lipid bilayer) bind to receptors in
the plasma membrane.
There are two subclasses of hydrophilic hormones:
(1) Peptide hormones such as insulin and glucagon
(2) Small charged molecules, often biogenic amines, such as
epinephrine (adrenalin) and histamine.
Transport
Signaling molecules/ hormone released into the circulation can circulate
either freely or bound to carrier proteins, also known as binding proteins.
The binding proteins serve as a reservoir for the hormone and prolong the
hormone's half-life, the time during which the concentration of a hormone
decreases to 50% of its initial concentration.
The free or unbound hormone is the active form of the hormone, which
binds to the specific hormone receptor. Thus, hormone binding to its carrier
protein serves to regulate the activity of the hormone by determining how
much hormone is free to exert a biologic action. Most carrier proteins are
globulins and are synthesized in the liver.
Target cell
Any cell in which the hormone (ligand) binds to its receptor, whether or not a
biochemical or physiologic response has yet been determined. Several factors
determine the response of a target cell to a hormone.
1. Determinants of the Concentration of a Hormone at the Target Cell
 The rate of synthesis and secretion of the hormones.
 The proximity of the target cell to the hormone source (dilution effect).
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 The dissociation constants of the hormone with specific plasma transport

proteins (if any).
 The conversion of inactive or sub-optimally active forms of the hormone
into the fully active form.
 The rate of clearance from plasma by other tissues or by digestion,
metabolism, or excretion.
2. Determinants of the Target Cell Response
 The number, relative activity, and state of occupancy of the specific
receptors on the plasma membrane or in the cytoplasm or nucleus.
 The metabolism (activation or inactivation) of the hormone in the target
cell.
 The presence of other factors within the cell that are necessary for the
hormone response.
 Up- or down-regulation of the receptor consequent to the interaction
with the ligand.
 Post-receptor desensitization of the cell, including down-regulation of
the receptor.

Hormone Receptor
A target cell is defined by its ability to selectively bind a given hormone to
its cognate receptor.
Several biochemical features of this interaction are important in order for
hormone-receptor interactions to be physiologically relevant:
 Binding should be specific, i.e, displaceable by agonist or antagonist;
 Binding should be saturable;
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 Binding should occur within the concentration range of the expected

biologic response.
All receptors are proteins and have at least two functional domains.
 A hormone recognition domain binds the hormone ligand
 A second region generates a signal that couples hormone recognition
to some intracellular function.
The dual functions of binding and coupling ultimately define a receptor, and
it is the coupling of hormone binding to signal transduction—so-called
receptor-effector coupling—that provides the first step in amplification of the
hormonal response. This dual purpose also distinguishes the target cell
receptor from the plasma carrier proteins that bind hormone but do not
generate a signal.
This coupling results in generation of an intracellular signal that can either:
 Regulate the activity of a selected set of genes, thereby altering the
amount of certain proteins in the target cell.
 Or affect the activity of already existing specific proteins, including
enzymes and transporter or channel proteins.
Types of receptors
Intracellular receptors
 Nuclear receptors
 Cytoplasmic receptors
Cell membrane receptors
 Ion channel receptors
 Receptors that are kinases or bind kinases
 Receptors that work through second messengers
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Ion channel receptors

Here binding of the ligand again causes a conformational change in the

protein but this time such that a specific ion channel is opened. This allows a
certain ion to flow through that subsequently alters the electric potential
across the membrane. For example, at the nerve–muscle junction the
neurotransmitter acetylcholine binds to specific receptors that allow Na+ ions
to flow into and K+ ions out of the target cell
Receptors that are kinases or bind kinases
On binding of the ligand to its extracellular face, the cell-surface receptor
undergoes a conformational change and activates an intrinsic enzyme activity.
Example, insulin receptor.

Dr Saira Saad
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Receptors that work through second messengers

These receptors, known as G protein-coupled receptors (GPCR), are

characterized by an extracellular ligand-binding region, seven transmembrane
helices, and an intracellular domain that interacts with G proteins. It is also
called heptahellicular receptor.
G proteins consist of three subunits; α,β and γ.The α subunit binds GTP or
GDP. The β and γ subunits do not bind nucleotides.
a. The hormone-receptor complex catalyzes the exchange of GDP for GTP
by the G protein. The receptor alone does not catalyze this exchange.

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b. When GDP is exchanged for GTP on the α subunit Gα-GTP dissociates

from Gβγ. Gα-GTP is the active form.
The effect of the active form (Gα-GTP) depends upon the specific type of G
protein. There are several different types of G proteins:
 Gs stimulates the enzymes Adenylate cyclase
 Gi inhibits the enzyme adenylate cyclase
 Gq stimulates the enzyme phospholipase C.
Function: The Gα subunit of all G proteins is a GTPase. It slowly
hydrolyzes its bound GTP to GDP and thereby returns to its inactive, GDP
bound state. Gα then reassociates with Gβγ, where it remains until it is
reactivated by a hormone- receptor complex.
Clinical Notes:
Cholera toxin is an enzyme produced by bacterium vibrio cholerae
Action: Cholera toxin modifies the α subunits of Gs which blocks the hydrolysis
of GTP to GDP. This prevents the inactivation of Gs by this mechanism.
Result: Persistently high level of cyclic AMP which causes the epithelial cells of
the intestine to transport Na+ and water into the intestinal lumen which results
in severe diarrhea.

Pertussis toxin is produced by Bordetella pertussis the bacterium that causes

whooping cough.
Action: It modifies the α subunit of Gi.
Result: The modification prevents Gi from exchanging GDP to GTP therefore
the modified Gi protein is unable to block the activation of adenylate cyclase.

ADENYLYL CYCLASE
This is a membrane-bound enzyme that converts ATP to 3',5'-adenosine
monophosphate (also called cyclic AMP or cAMP). The chemical signals are
most often hormones or neurotransmitters, each of which binds to a unique

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type of membrane receptor. Therefore, tissues that respond to more than one
chemical signal must have several different receptors, each of which can be
linked to adenylyl cyclase .
SECOND MESSENGERS
• cAMP
• cGMP
• PIP2
• Ca – Calmodulin

Hormones that act thru cAMP:

• α2-Adrenergic catecholamines
• β-Adrenergic catecholamines
• Adrenocorticotropic hormone
• Antidiuretic hormone
• Calcitonin
• Glucagon
• Parathyroid hormone

MOA of cAMP
A hormone binds to G-protein
coupled receptor. Α subunit of G-protein
activates adenylate cyclase which in turn
coverts ATP to cAMP. Next cAMP
activates family of enzymes called cAMP-
independent protein kinases , for
example, protein kinase A. Cyclic AMP
activates protein kinase A by binding to
its two regulatory subunits, causing the
release of active catalytic subunits. The
active subunits catalyze the transfer of
phosphate from ATP to specific serine or
threonine residues of protein substrates.
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The phosphorylated proteins may act directly on the cell’s ion channels, or, if
enzymes, may become activated or inhibited. Protein kinase A can also
phosphorylate proteins that bind to DNA, causing changes in gene expression.
The phosphate groups added to proteins by protein kinases are removed
by protein phosphatases —enzymes that hydrolytically cleave phosphate
esters. This ensures that changes in protein activity induced by
phosphorylation are not permanent.
cAMP is rapidly hydrolyzed to 5'-AMP by cAMP phospho diesterase , one
of a family of enzymes that cleave the cyclic 3',5'-phosphodiester bond. 5 '-
AMP is not an intracellular signaling molecule. Thus, the effects of
neurotransmitter- or hormone-mediated increases of cAMP are rapidly
terminated if the extracellular signal is removed. Phosphodiesterase is
inhibited by methyl xanthine derivatives, such as theophylline and caffeine.

Dr Saira Saad
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Hormones that act thru cGMP:

• Atrial natriuretic factor
It activates cGMP and causes:
-Natriuresis
-Dieresis
-Vasodilation
-Inhibition of Aldosterone secretion

• Nitric oxide
Activate cGMP dependant protein kinase. Causes phosphorylation of a
number of smooth muscle proteins resulting in smooth muscle relaxation
and vasodilation.

Hormones that act thru Phosphatidyl inositol bisphosphate (PIP2):

• Angiotensin II • Acetylcholine
• Antidiuretic hormone • α1-Adrenergic catecholamines
(vasopressin)
• Cholecystokinin • Oxytocin
• Gastrin • Platelet-derived growth factor
• Gonadotropin-releasing hormone • Substance P

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The phosphorylation of membrane-bound phosphatidylinositol produces

polyphosphoinositides, for example, phosphatidylinositol 4,5 bisphosphate
(PIP2). The degradation of PIP2 by phospholipase C occurs in response to the
binding of a variety of neurotransmitters, hormones, and growth factors to
receptors on the cell membrane. The products of this degradation, inositol
1,4,5-trisphosphate (IP3) and diacylglycerol (DAG), mediate the mobilization of
intracellular calcium and the activation of protein kinase C , respectively, which
act synergistically to evoke specific cellular responses.
Proteins regulated by Ca -Calmodulin
• Adenylyl cyclase
• Ca ⁺⁺ -dependant kinases
• Some cytoskeletal proteins
• Some ion channels (calcium channels type –I)
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TYROSINE KINASE PATHWAY

 Receptors with intrinsic/ integral tyrosine kinase
activity.
• Insulin , Epidermal growth factor

Stimulation of the intrinsic tyrosine kinase activity of the insulin receptor

marks the initial event, resulting in increased tyrosine phosphorylation of the
receptor and then one or more of the insulin receptor substrate molecules (IRS
1–4). This increase in phosphotyrosine stimulates the activity of many
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intracellular molecules such as GTPases, protein kinases, and lipid kinases, all of
which play a role in certain metabolic actions of insulin.

 Receptors that bind cytoplasmic proteins with tyrosine kinase activity.

JAK- STAT pathway:
• Growth hormone, Prolactin & Erythropoietin

The hormone–receptor interaction promotes binding and activation of

cytoplasmic protein tyrosine kinases, such as Tyk-2, Jak1, or Jak2.
These kinases phosphorylate one or more cytoplasmic proteins, which
then associate with other docking proteins through binding to SH2 domains.
One such interaction results in the activation of a family of cytosolic proteins
called
signal transducers and activators of transcription (STATs).
The phosphorylated STAT protein dimerizes and translocates into the
nucleus, binds to a specific DNA element such as the interferon response
element, and activates transcription.

Dr Saira Saad
 CELL SIGNALLING

NF-κB pathway

Activation of NF-κB pathway regulates transcription of the multitude of

genes important for inflammatory response.
Glucocorticoid hormones are therapeutically useful agents for the treatment of
a variety of inflammatory and immune diseases by inhibiting NF-κB pathway.
(1) Glucocorticoids increase IκB mRNA, which leads to an increase of IκB
protein and more efficient sequestration of NF-κB in the cytoplasm.
(2) The glucocorticoid receptor competes with NF-κB for binding to
coactivators.
(3) The glucocorticoid receptor directly binds to the p65 subunit of NF-κB
and inhibits its activation.

LIPOPHILIC HORMONES MOA:

The lipophilic group I hormones diffuse through the plasma membrane of
all cells but only encounter their specific, high affinity intracellular receptors in
target cells. These receptors can be located in the cytoplasm or in the nucleus
of target cells.
The hormone–receptor complex first undergoes an activation reaction. The
activated receptor moves into the nucleus and binds with high affinity to a
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specific DNA sequence called the hormone response element (HRE). The DNA-
bound, liganded receptor serves as a high-affinity binding site for one or more
coactivator proteins, and accelerated gene transcription typically ensues when
this occurs.
Certain hormones (thyroid hormones) diffuse from the extracellular fluid
across the plasma membrane and go directly into the nucleus. In this case, the
cognate receptor is already bound to the HRE. However, this DNA-bound
receptor fails to activate transcription because it exists in complex with a
corepressor.
The association of ligand with these receptors results in dissociation of the
corepressor(s). The liganded receptor is now capable of binding one or more
coactivators with high affinity, resulting in activation of gene transcription.
By selectively affecting gene transcription and the consequent production
of appropriate target mRNAs, the amounts of specific proteins are changed and
metabolic processes are influenced.
 Glucocorticoids  Estrogens
 Mineralocorticoids  Progestins
 Androgens  Calcitriol
 Retinoic acid  Thyroid hormones

Dr Saira Saad