Cell Signalling

Need for cell signalling

• All forms of cells rely on communication with their surrounding environment.
• This communication is needed to adjust their internal environment according to the situation
outside.

• Examples of such adjustments can be

1. Movement towards or away from the food.
2. Movement towards or away from chemical gradient.
3. Responding to solute concentration changes in the medium.
4. Responding to growth factors.
5. Responding to hormones.
6. Responding to voltage changes.
7. Responding to insults to the cell.

And many more such examples.

• Now, the question is how do they respond to the changes in the surrounding medium??
• The answer is cell signalling.
• Basic outline of how cell signalling works:

• It has following basic steps in all its forms

1. An extracellular signal in the form of a chemical molecule, generally called as ligand.

2. Ligand binds to its receptor on the target cell. Usually, the receptor is a transmembrane
protein but sometimes it can be a soluble protein found in cytosol or nucleus.

3. This receptor-ligand interaction conveys the outer situation inside the cell and initiates a
series of reactions.

4. This series of reactions is focussed on two things in most cases

1. Signal amplification by generating the second messenger.
2. Activating the genes and proteins that are needed for appropriate response to the signal.

5. Now cell begins to show its response to the signalling ligand.

E.g. glycogenesis in response to insulin signalling.
Features of signal transducing systems
 Types of signalling
• Based on the distance the ligand molecule has to travel signalling is of following types
1. Autocrine signalling: The ligand molecule is diffusible but binds to the same cell that has secreted it. E.g.
cytokine signalling in immune system.

2. Paracrine signalling: The ligand molecule is diffusible and binds to the neighbouring cells that are present
around the secreting cell. E.g. Prostaglandin signalling during inflammation.
3. Endocrine signalling: The ligand has to travel a long distance in blood to reach its target
cell. E.g. Hormone signalling

Juxtacrine signalling

4. Juxtacrine signalling: In this ligand is non-diffusible. Thus, receptor and ligand both are
fixed transmembrane molecules and found on adjacent cells that are in touch with each
other.
E.g. Delta-notch signalling during embryonic development.
Types of signal transducing systems based on receptors
 Receptor enzymes such as insulin receptor
• General discussion:These proteins have a ligand-binding domain on the extracellular surface of
the plasma membrane and an enzyme active site on the cytosolic side,

• These two domains are connected by a single transmembrane segment.

• Commonly, the receptor enzyme is a protein kinase that phosphorylates Tyr residues in specific
target proteins; the insulin receptor is the prototype for this group.

• In plants, the protein kinase of receptors is specific for Ser or Thr residues.

• Other receptor enzymes synthesize the intracellular second messenger cGMP in response to
extracellular signals. The receptor for atrial natriuretic factor is typical of this type.
1. The receptor is a monomer in
absence of ligand.

2. On binding with ligand it

undergoes dimerization to form a
homodimer.

3. The tyrosine kinase activity in the

cytosolic domain begins to
phosphorylate the tyrosine residue
in its partner. This process is called
as cross phosphorylation.

4. After this begins the downstream

signalling in the form of a cascade.
Insulin signalling, its basic steps and effects
Effect of insulin on muscle cells
Variation of RTK
Receptor enzymes with guanylyl cyclase activity

Effects of cGMP are mediated by activating the enzyme

Protein kinase G or PKG. It causes phosphorylation of
Ser or Thr.
Termination of cGMP signalling
 Nitric oxide signalling and its effects

Nitric oxide production How nitroglycerine works??

Nitric oxide, Sildenafil citrate and Nobel prize in medicine
 G-protein coupled receptors(GPCR) and their second messenger
• They are called as serpentine receptor because they have 7 transmembrane domains that
resemble the twists of snake.

• They are called as G-protein coupled because they have a cytosolic region that is found
associated with a guanosine-nucleotide binding protein or G-protein.
• Epinephrine receptors are called as adrenergic receptors and are of 4 general types viz α1,
α2, β1, β2.
• The β family of receptors are found in the muscles, liver and adipose tissues.

Epinephrine signalling G-protein cycle

 Why we call cAMP as second messenger??

Adenylyl cyclase
 Termination of epinephrine signalling

Cyclic nucleotide PDE

 Modulations of cAMP during signal transduction
• Not just epinephrine but many other hormones use cAMP as their second messenger such as
glucagon, ACTH and some prostaglandins etc.

• All of them act to temporarily increase the concentration of cAMP in the cell. However, it is not
the only modulation possible on cAMP.

• Some molecules act to decrease the levels of cAMP in the cell.

• They do so by inhibition of the enzyme adenylyl cyclase.

• Examples of such molecules include epinephrine acting through acting through α2 receptors,
somatostatins.
• Receptors for these proteins are linked with
inhibitory G-proteins or Gi.

• They are structural homologs of Gs proteins.

• Once activated these proteins bind to GTP and

act to inhibit the enzyme adenylyl cyclase.
 Disruption of G-protein signalling and
toxins
• Cholera toxin: It is responsible for the symptoms of cholera and is an enzyme.

• Once inside the cell in small intestine, it causes ADP-ribosylation of the α-subunit of the Gs
protein.
• The ADP-ribose is taken from NAD+ .

• This reaction blocks the GTPase activity of the Gα protein.

• Thus, adenylyl cyclase is permanently on and continues to produce cAMP at high levels.

• As a result, chloride channels are left open and this leads to loss of electrolytes.
• Pertussis toxin: It is responsible for symptoms of whooping cough and is an enzyme like
the cholera toxin.
• It also acts by causing the ADP-ribosylation reaction by transferring the ADP-ribose
moiety from NAD+.

• However, it does so on the α-subunit of the Gi protein.

• This reactions blocks the displacement of GDP by GTP.

• Thus, it fails to inhibit the adenylyl cyclase enzyme and cAMP production continues.
• According to the classical A-F system, GPCRs can be grouped into 6 classes based on sequence
homology and functional similarity:[
• Class A (or 1) (Rhodopsin-like)
• Class B (or 2) (Secretin receptor family)
• Class C (or 3) (Metabotropic glutamate/pheromone)
• Class D (or 4) (Fungal mating pheromone receptors)
• Class E (or 5) (Cyclic AMP receptors)
• Class F (or 6) (Frizzled/Smoothened)

• More recently, an alternative classification system called

• GRAFS (Glutamate, Rhodopsin, Adhesion, Frizzled/Taste2, Secretin) has been proposed for
vertebrate GPCRs.
• They correspond to classical classes C, A, B2, F, and B
Other secondary messengers
Signalling by steroid hormones
Plasma membrane receptors of Progesterone
Bacterial two component system
Assignment: Cell signalling
• Q.1 Which of the following statements is FALSE for the nitric oxide gas?

(a) An intracellular signaling molecule

(b) Deamination of histidine results into nitric oxide production

(c) Stimulates guanylyl cyclase to produce cGMP

(d) Can be produced by activated neutrophils

• Q.2 G-proteins are involved in relaying signals through G-protein linked
receptors. Which of the following forms of G-protein is considered to be in
active state?

(a) G-protein - ADP

(b) G-protein – ATP

(c) G-protein – GDP

(d) G-protein - GTP

• Q.3 The ‘A’ fragment of diphtheria toxin catalyzes the addition of an ADP-
ribose group to the eukaryotic elongation factor EF-2.
The source for the ADP-ribose group is

(a) FAD

(b) FMN

(c) NAD+

(d) NADP+
• Q.4
• Q.5 Cholera toxin manifests its action by
P. the ADP-ribolysation of Gs protein

Q. the transfer of ADP-ribose from NAD+ to the Gi protein

R. the inhibition of phosphodiesterase

S. the activation of adenylate cyclase

(a) P and R
(b) P and S
(c) Q and S
(d) Q and R
• Q.6 Which one of the following second messengers targets protein kinase A?

(a) cAMP

(b) cGMP

(c) Diacylglycerol

(d) Inositol 1, 4, 5-triphosphate

• Q.7 The INCORRECT statement regarding second messenger, adenosine
3`,5` -cyclic nucleotide monophosphate (cAMP), is

(a) it acts as a second messenger for many regulatory molecules

(b) it acts as an intracellular second messenger in neurons

(c) it activates specific cyclic nucleotide dependent protein kinases

(d) it provides source of energy for cells

• Q.8 The preferred ligand for SH2 domain is

(a) serine-phosphorylated peptide

(b) tyrosine-phosphorylated peptide

(c) glucose-6-Phosphate

(d) cyclic AMP

• Q.9 The binding of a hormone to its receptor activates adenylyl cyclase
through a stimulatory G protein. If, due to a mutation, the G-protein binds
but does NOT hydrolyze, GTP, the consequence will be

(a) Adenylyl cyclase will be continuously activated

(b) Adenylyl cyclase will never be activated

(c) Adenylyl cyclase will be occasionally activated

(d) Adenylyl cyclase will be degraded

• Q.10 Signaling pathways usually comprise of several intermediate steps that
are arranged in the form of a cascade. What is the primary outcome of such
an arrangement?

(a) Specificity of signal transduction

(b) Specificity of the cellular response

(c) Amplification of the cellular response

(d) Fine-tuning of the cellular response

• Q.11 Which of the following statements is/are CORRECT for G protein-
coupled receptor (GPCR) mediated signaling?

(a) GPCRs contain seven membrane spanning regions.

(b) GPCRs are linked to heterotrimeric G protein consisting of α,β and γ

subunits.
(c) In the absence of GPCR interacting ligand, a subunit of G protein is bound
to GTP and complexed with βγ subunits.

d) In the presence of GPCR interacting ligand, GTP is displaced from α

subunit of G protein by GDP, GDP bound α subunit dissociates from βγ dimer
and activates the effector
• Q.12 The intracellular messengers formed by the activation of
phosphoinositide cascade are

(a) phoshophatidylinositol-4,5-bisphosphate

(b) inositol-1,4,5-triphosphate

(c) diacylglycerol

(d) inositol-4-phosphate
• Q.13 Which of the following statements are TRUE for phosphoinositide
signaling cascade?

(a) Phospholipase A catalyzes cleavage of PIP2

(b) Generation of IP3 transiently increases cytosolic Ca+2 concentration

(c) Ca+2 facilitates the activation of protein kinase C

(d) DAG always activates protein kinase A

• Q.14 Cell type A secretes molecule X into the culture medium. Cell type B in
the same culture responds to the molecule X by expressing protein Y. Which
one of the following modes of signaling represents the interaction between A
and B?

(a) Autocrine

(b) Juxtacrine

(c) Paracrine

(d) Intracrine
• Q.15 Highly developed organisms are made up of many different cell types. In
order for the organism to function properly, the cells must work together. This
is accomplished by various types of chemicals that act as messengers to the
cells. Each chemical messenger has a specific effect on its target cell. This is
called signal transduction. Hormones are good examples of chemical
messengers. Steroid hormones such as estrogen are non-polar and can thus
diffuse across the cell membrane. However, polar hormones cannot. For their
message to enter the cell these polar hormones must

(a) form a micelle

(b) bind to receptors on the cell surface

(c) bind to g proteins

• Q.16 Binding of erythropoietin to its EPO receptor leads to activation of
signaling pathway termed as
(a) JAK STAT pathway

(b) NF kβ pathway

(c) Apaf smad pathway

(d) Tyrosine kinase pathway

• Q.17 Which one of the following statements about receptor-enzyme is FALSE?

(a) A receptor - enzyme has an extracellular ligand binding domain, a transmembrane

domain and an intracellular catalytic (enzyme) domain.

(b) Many types of receptor enzymes are found in animals.

(c) The signal transduction pathways of receptor - enzyme involve phosphorylation

cascades.

(d) Receptor - enzymes interact directly with intracellular G-proteins.

• Q.18 Ras contributes to cancer due to its

(a) innate GTPase activity

(b) promoter DNA binding activity

(c) serine / threonine kinase activity

(d) tyrosine phosphorylation activity