RECEPTORS

Receptors are protenious molecules/ elements present in the cell wall. These are acts as sensitizing
elements of chemical communication system that coordinates the functions of different cells in the body, on
which endogenous messengers like hormones, neurotransmitters and other mediators such as cytokines and
other factors act to bring about desired biological effect.

Types of receptors: Based on molecular structure and nature of the linkage (transduction mechanism) we can
distinguish the receptors in to four types.

1. Ligand-gated ion channels

2. G-protein coupled receptors
3. Kinase-linked receptors
4. Nuclear receptors

Type-I: Ligand gated ion channels (ionotropic receptors)

Molecular structure:

The ligand gated ion channels have structural features in common with other ion channels. The nicotinic
cholinergic (Ach) receptor was first cloned and it composed of 5 subunits (2+ + + ) enclosing a cylindrical ion
channel. The oligomeric structure possesses two Ach binding sites, each laying the interface between one of the
two subunits and its neighbor. Both must bind Ach molecule in order to activate the receptor. The receptor is
usually a pentameric protein; all subunits in addition to large intra and extracellular segments, generally have
four membrane spanning domains(sites) in each of which the amino acid chain traverses the width of the
arranged round the channel like a rosette and usually subunit bear the agonist binding site.
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The Gating mechanism:

Receptors of this type control fastest synaptic events in the nervous system, in which the neurotransmitter act on
post synaptic membrane of a nerve and transiently increase the permeability of particular ions. Most excitatory
neurotransmitters such as Ach in neuromuscular junction and glutamate in the CNS act by increase Na + and K+
permeability. The action o transmitter reaches to a peak in a fraction of millisecond and usually within a few
milliseconds.

(A) Normally the channel is closed. (B) When two molecules of acetylcholine bind to the two subunits.

Ex: nicotinic Ach receptors, GABA A receptors glutamate (NMDA, AMPA) receptors
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Type II: G-Protein Coupled Receptors (metabotropic receptors)

Depending on the length of extracellular loop the G-Protein coupled receptors are classified into 3 families.

Molecular structure:
These are a large family of cell membrane receptors which are linked to the effector (enzyme/ channel/ carrier
protein) through one or more GTP-activated proteins (G-proteins) for response effectuation. The molecule has 7
-helical membrane spanning hydrophobic amino acid (AA) segments which run into 3 extracellular and 3
intracellular loops. The agonist binding site is located somewhere between the helices on the extracellular face,
while another recognition site formed by cytosolic segments binds the coupling G-protein. The G-proteins float
in the membrane with their exposed domain lying in the cytosol, and are heterotrimeric in composition (, and
y subunits), the subunit possessing GTPase activity.

Ex: Muscarinic Ach receptors; adrenergic receptors; H2, 5-HT, opiate, peptide hormone receptors
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Function of G-Protein:

In the inactive state GDP is bound to their exposed domain; activation through the coupling of the -subunit to
an agonist-occupied receptor causes the displacement of GDP by intracellular GTP; the -GTP complex then
dissociates from the receptor and from the complex, and activates/ inhibits the effector protein (target 1,
which may be an enzyme, such as adenylate cyclase, or an ion channel). The GTPase activity of the -subunit is
increased when the target protein is bound, leading to hydrolysis of the bound GTP to GDP, whereupon the -
subunit reunites with . The complex may also activate a target protein (target 2). The , subunits have also
been shown to modulate certain effectors like receptor operated K+ channels, adenylylcyclase (A C) and
phospholipase C.
The main G-protein subtypes and their functions
Subtypes Associated receptors Main effectors Notes
G
subunits
Gs Many amine and other Stimulates adenylyl cyclase, causing Activated by cholera toxin,which blocks GTPase
receptors (catecholamines, increased cAMP formation. activity, thus preventing inactivation.
histamine, serotonin)
Gi As for Gs, also opioid, Inhibits adenylyl cyclase, decreasing Blocked by pertussis toxin, which prevents
cannabinoid receptors cAMP formation. dissociation of complex.
Go As for Gs, also Limited effects of subunit (effects Blocked by pertussis toxin. Occurs mainly in
opioid,cannabinoid receptors mainly due to -subunits) nervous system.
Gq Amine, peptide and prostanoid Activates phospholipase C, increasing
receptors production of second messengers
-------
inositol trisphosphate
&diacylglycerol.
G All GPCRs activate potassium channels Many G isoforms identified, but specific
subunits inhibit voltage-gated calcium functions are not yet known. G-mediated
channels effects probably require higher levels of GPCR
activate GPCR kinases (p. 40) activation than G-mediated effects.
activate mitogen-activated protein
kinase cascade.
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TARGETS FOR G-PROTEINS:

The main targets for G-proteins, through which GPCRs control different aspects of cell function

Adenylyl cyclase, the enzyme responsible for CAMP formation

Phospholipase c, the enzyme responsible for inositol phosphate(IP3) and diacylglycerol (DAG) formation

Ion channels, particularly calcium and potassium channels

Adenylyl cyclase (CAMP Pathway):

Adrenaline (Adr) binds to -adrenergic receptor (-R) on the cell surface inducing a conformational change which
permits interaction of the G-protein binding site with the stimulatory G-protein (Gs). The activated Gs now binds
GTP causing its active subunit to dissociate and in turn activate the enzyme adenylyl cyclase (AC) located on the
Cytosolic side of the membrane: A TP is hydrolysed to cAMP which phosphorylates and thus activates cAMP
dependent protein kinase (PKA) The PKA phosphorylates many functional proteins including troponin and
phospholamban, so that they interact with Ca 2+, respectively resulting in increased force of contraction and faster
relaxation. Calcium is made available by entry from outside (direct activation of myocardial membrane Ca 2+
channels by Gs and through their phosphorylation by PKA) as well as from intracellular stores. Action of
Acetylcholine (ACh) on muscarinic M2 receptor (M2-R), also located in the myocardial membrane, can similarly
activated an inhibitory G-protein (Gi) which then opposes the activation of AC by Gs.
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IP3 and DAG Pathway:

The agonist, e.g. histamine binds to its H, receptor (H1-R) and activates the G-protein (Gq). which inturn activates
membrane bound phospholipase C (Plc) that hydrolyses phosphatidyl inositol 4, 5-bisphosphate (PIP2), a
membrane bound phospholipid. The products inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG) act as
second messengers. The primary action of IP3 is facilitation of Ca2+ mobilization from intracellular organellar
pools, while DAG in conjunction with Ca2+ activates protein kinase C (PKc) which phosphorylates and alters the
activity of a number of functional and structural proteins. Cytosolic Ca2+ is a veritable messenger: combines with
calmodulin (CAM) to activate myosin light chain kinase (MLCK) inducing contraction, and another important
regulator calcium-calmodulin protein kinase (CCPK) Several other effectors are regulated by Ca2+ in a CAM
dependent or independent manner.

Channel regulation: activation of G-Protein can also open/close ionic channels specific for Ca2+, K+/Na+ to ring
depolarization or repolarization.
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Type-III: Kinase Linked and Related Receptors

These are large heterogenous group of membrane receptors responding to Protein mediators. They comprise an
extracellular ligand-binding domain linked to an intracellular domain by a single transmembrane helix.
Ex-Growth factors,Insulin,Cytokine receptors(having protein/tyrosine kinase activity).
The receptors for Atrial natriuetic factor (Guanylate cyclase type). In case of insulin receptor the extra cellular
domain consists of a seperte poly peptide which is linked to the rest of the molecule by di-sulphide bonds. The
growth factor receptor consists of a single long chain of over 1000 residues. Cytokine receptors are similar but
are often dimeric. The growth factor and insulin (intrinsic enzyme receptors) receptor in the intra cellular region
possess Tyrosine Kinase activity and incorporates both ATP and substrate binding sites. Cytokine receptors do not
have intrinsic Kinase activity/ catalytic domain, but associates when activated by ligand binding with JAK (JAK-
STAT Kinase binding receptors).

MECHANISM
The tyrosine kinase receptors signal transduction generally begins with the binding of ligand to the receptor
extracellular site. The resulting conformational changes cause in receptors molecule involves dimerization of
receptors, followed by autophosphorylation of Tyrosine residue. The phosphor Thyrosin residue acts as receptors
for the SH2 domain of a variety of intra cellular proteins, thereby allowing control of many cell functions.
There are two signal transduction path ways
A. Ras/Raf / Map kinase path way- The intracellular domain is either a protein kinase or guanyl cyclase
B. Jak/Stat pathway-These receptors (cytokine) do not having any intrinsic activity/ catalytic doamin

A. Ras/Raf path way:

It mediates the effect of many growth factors and Mitogens. Ras-proto-oncogen product and function coneys
signals from SH2 binding protein Grb which phospho related by recepotrs tyrosin kinase.Activation of Ras turns
to activation of Raf which is the 1 st sequences of serine by Theronine residues and phosphorylates each of which
activates the next step.The last step in activation of Map (Mitogen-activated Protein) kinase, Phosphorylates one
or more transcription factors and initiates gene expression resulting in a variety of cell response including cell
division.(Growth and Differentiation)
B. Jak/Stat pathway:
Dimeraization of thses receptors when cytokinin binds and this attracts a tyrosine kinase unit (JAK) to associate
with and phosphorylate receptor dimer. Tragets for phosphorylation of JAKs are family of transcription
factors(STATS). These are SH2 doamin proteins that bind to phosphotyrosine group of receptor-JAK complex, and
themselves phosphorylated. Thus activated STAT migartes to the nucleus and activates gene expression.
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Type-IV: Nuclear Receptors (regulates gene transcription)

Molecular structure

Most receptors located in nucleus and ligands are liphophillic compounds that can readily cross the cell
membrane. It pocess highly conserved area of about 60 residues in the middle of the molecule constitutes of
DNA binding domain of the receptor. It contain two loops of about 15 residues each knotted together by a
cluster of four cysteine residues surrounding a zinc atom; these structures occur in many proteins that regulate
DNA transcription, and the fingers are believed to wrap around the DNA helix. The hormone-binding domain lies
downstream of this central region, while upstream lies a variable region that is responsible for controlling gene
transcription On binding a steroid molecule, the receptor changes its conformation, which facilitates the
formation of receptor dimers. These dimers bind to specific sequences of the nuclear DNA, known as
hormone-responsive elements, which lie about 200 base pairs upstream from the genes that are
regulated.

Mechanism of action:

The glucocorticoid (G) penetrates the cell membrane and binds to the glucocorticoid receptor (GR) protein that
normally resides in the cytoplasm. The G R has a steroid binding domain near the carboxyl terminus and a mid
region DNA binding doma1n having two Zinc lingers', each made up of a loop of amino acids with chelated zinc
ion Binding of the steroid to GR dissociates the complexed proteins removing their inhib1tory influence on
dimerization region that overlaps the steroid binding domain Is exposed, promoting dimerization of the occupied
receptor The steroid bound receptor diamer translocates on the nucleus and interacts with specific DNA
sequences called 'glucocorticoid responsive elements' (GREs) within the regulatory region of appropriate genes
The expression of these genes is consequently altered resulting in promotion (or expression) of their
transcription Tile specific m RNA thus produced is directed to the ribosome where the message i s translated
into a specific pattern of protein synthesis, which in turn modifies cell function.