RECEPTORS

Naresh Panigrahi
Assistant professor
GITAM INSTITUTE OF PHARMACY
 TARGETS FOR DRUG ACTION
• The protein targets for drug action on
mammalian cells that are described can be
broadly divided into:
• receptors
• ion channels
• enzymes
• carrier molecules (transporters).
 TYPES OF RECEPTORS
• Type 1: ligand-gated ion channels (also known as ionotropic
receptors).
• These are membrane proteins with a similar structure to other ion channels,
and incorporate a ligand-binding (receptor) site, usually in the extracellular
domain. Typically, these are the receptors on which fast neurotransmitters act.
• Examples include the nicotinic acetylcholine receptor ; GABA A receptor ; and
glutamate receptors of the NMDA, AMPA and kainate types
• Type 2: G-protein-coupled receptors (GPCRs).
• These are also known as metabotropic receptors or 7-transmembrane-
spanning (heptahelical) receptors. They are membrane receptors that are
coupled to intracellular effector systems via a G-protein (see below). They
constitute the largest family,5 and include receptors for many hormones and
slow transmitters,
• for example the muscarinic acetylcholine receptor, adrenergic receptors and
chemokine receptors.
• Type 3: kinase-linked and related receptors.
• This is a large and heterogeneous group of membrane receptors
responding mainly to protein mediators. They comprise an extracellular
ligand-binding domain linked to an intracellular domain by a single
transmembrane helix.
• In many cases, the intracellular domain is enzymic in nature (with
protein kinase or guanylyl cyclase activity).
• Type 3 receptors include those for insulin and for various cytokines and
growth factors the receptor for atrial natriuretic factor (ANF, is the main
example of the guanylyl cyclase type. The two kinds are very similar
structurally, even though their transduction mechanisms differ.
• Type 4: nuclear receptors.
• These are receptors that regulate gene transcription. The term nuclear
receptors is something of a misnomer, because some are actually
located in the cytosol and migrate to the nuclear compartment when a
ligand is present.
• They include receptors for steroid hormones thyroid hormone and
other agents such as retinoic acid and vitamin D.
 Type 1: ligand-gated ion Type 2: G-protein- Type 3: receptor Type 4: nuclear
channels coupled receptors kinases receptors

Location Membrane Membrane Membrane Intracellular

Effector Ion channel Channel or enzyme Protein kinases Gene transcription

Coupling Direct G-protein Direct Via DNA

Examples Nicotinic acetylcholine Muscarinic Insulin, growth Steroid receptors

receptor, GABA A receptor acetylcholine receptor, factors, cytokine
adrenoceptors receptors

Structure Oligomeric assembly of Monomeric dimericor Single transmembrane Monomeric structure

subunits surrounding structure comprising helix linking with separate
central pore seven transmembrane extracellular receptor receptor- and DNA-
helices domain to binding domains
intracellular kinase
domain
 TYPE 1: LIGAND-GATED ION CHANNELS
MOLECULAR STRUCTURE
• These molecules have structural features in common
with other ion channels.
• It is assembled from four different types of subunit,
termed α, β, γ and δ, each of Mr 40-58 kDa.
• The pentameric structure (α2, β, γ, δ) possesses two
acetylcholine binding sites, each lying at the interface
between one of the two α subunits and its neighbour.
• Both must bind acetylcholine molecules in order for
the receptor to be activated.
• The two acetylcholine-binding sites lie on the
extracellular parts of the two α subunits.
• One of the transmembrane helices (M2) from
each of the five subunits forms the lining of the
ion channel .
• The five M2 helices that form the pore are sharply
kinked inwards halfway through the membrane,
forming a constriction.
• When acetylcholine molecules bind, the α
subunits twist, causing the kinked M2 segments to
swivel out of the way, thus opening the channel
• Ligand-gated ion channels
• These are sometimes called ionotropic receptors.
• They are involved mainly in fast synaptic
transmission.
• There are several structural families, the commonest
being heteromeric assemblies of four or five
subunits, with transmembrane helices arranged
around a central aqueous channel.
• Ligand binding and channel opening occur on a
millisecond timescale.
• Examples include the nicotinic acetylcholine,
• GABA type A (GABAA), and
• 5-hydroxytryptamine type 3 (5-HT3) receptors.
TYPE 2: G-PROTEIN-COUPLED RECEPTORS

• The G-protein coupled receptors (GPCRs) is the largest of the four

superfamilies with some estimated 600 human receptor genes.
• Such as mAChRs,
• adrenoceptors,
• dopamine receptors,
• 5-HT receptors,
• opiate receptors,
• receptors for many peptides,
• purine receptors and many others,
• including the chemoreceptors involved in olfaction and
pheromone detection, and also many 'orphans‘.
• A large fraction of these are taste and odor sensing receptors.
 MOLECULAR STRUCTURE
• The receptors are also referred to as 7TM receptors due to the seven
α-helical transmembrane segments with an extracellular N-terminal
domain of varying length, and an intracellular C-terminal domain
found in all GPCRs.
• GPCRs are divided into three distinct families
• There is considerable sequence homology between the members of
one family,
• They share the same seven-helix (heptahelical) structure, but differ in
other respects, principally in the length of the extracellular N terminus
and the location of the agonist binding domain.
• Family A is by far the largest, comprising most monoamine,
neuropeptide and chemokine receptors.
• Family B includes receptors for some other peptides, such as
calcitonin and glucagon.
• Family C is the smallest, its main members being the metabotropic
glutamate and GABA receptors and the Ca2+-sensing receptors8.
• G-proteins comprise a family of membrane-
resident proteins whose function is to
recognise activated GPCRs and pass on the
message to the effector systems that generate
a cellular response.
• They are the go-between proteins, but were
actually called G-proteins because of their
interaction with the guanine nucleotides, GTP
and GDP.
• G-proteins consist of three subunits: α, β and γ .
• Guanine nucleotides bind to the α subunit, which has enzymic activity, catalysing
the conversion of GTP to GDP.
• The β and γ subunits remain together as a βγ complex.
• All three subunits are anchored to the membrane through a fatty acid chain,
coupled to the G-protein through a reaction known as prenylation.
• In the 'resting' state ,the G-protein exists as an unattached αβγ trimer, with GDP
occupying the site on the α subunit.
• When a GPCR is activated by an agonist molecule, a conformational change
occurs, involving the cytoplasmic domain of the receptor, causing it to acquire
high affinity for αβγ. Association of αβγ with the receptor causes the bound GDP
to dissociate and to be replaced with GTP (GDP-GTP exchange), which in turn
causes dissociation of the G-protein trimer, releasing α-GTP and βγ subunits;
these are the 'active' forms of the G-protein, which diffuse in the membrane and
can associate with various enzymes and ion channels, causing activation of the
target .
• Signalling is terminated when the hydrolysis of GTP to GDP occurs through the
GTPase activity of the α subunit. The resulting α-GDP then dissociates from the
effector, and reunites with βγ, completing the cycle.
 TYPES OF G-PROTEIN
• Four main classes of G-protein (Gs, Gi, Go and Gq) are of
pharmacological importance.
• They show selectivity with respect to both the receptors and
the effectors with which they couple, having specific
recognition domains in their structure complementary to
specific G-protein-binding domains in the receptor and
effector molecules.
• Gs and Gi produce, respectively, stimulation and inhibition of
the enzyme adenylyl cyclase .
• The G-proteins can be thought of as the intramembrane
managers, bustling between receptors and effectors,
controlling this microcosm but communicating very little with
the world outside.
Subtypes Associated receptors Main effectors Notes

G α subunits

Gαs Many amine and other receptors (e.g. Stimulates adenylyl Activated by cholera
catecholamines, histamine, serotonin) cyclase, causing increased toxin,which blocks GTPase
cAMP formation. activity, thus preventing
inactivation.

Gαi As for Gαs, also opioid, cannabinoid Inhibits adenylyl cyclase, Blocked by pertussis toxin,
receptors decreasing cAMP which prevents dissociation
formation. of αβγcomplex.

Gαo As for Gαs, also opioid,cannabinoid ?Limited effects of Blocked by pertussis toxin.
receptors αsubunit (effects mainly Occurs mainly in nervous
due to βγsubunits) system.

Gαq Amine, peptide and prostanoid Activates phospholipase C, -

receptors increasing production of
second messengers inositol
trisphosphate and
diacylglycerol (see p. 38).
Gβγ subunits All GPCRs Many Gβγisoforms
As for Gα subunits . Also: identified, but specific
• activate potassium functions are not yet
channels known. Gβγ-mediated
• inhibit voltage-gated effects probably require
calcium channels higher levels of GPCR
• activate GPCR kinases activation than Gα-
• activate mitogen- mediated effects.
activated protein kinase
cascade.
 TARGETS FOR G-PROTEINS
• The main targets for G-proteins, through which GPCRs
control different aspects of cell function are:
• Adenylyl cyclase, the enzyme responsible for cAMP
formation
• Phospholipase - C, the enzyme responsible for inositol
phosphate and diacylglycerol (DAG) formation
• ion channels, particularly calcium and potassium
channels
• Rho A/Rho kinase, a system that controls the activity of
many signalling pathways controlling cell growth and
proliferation, smooth muscle contraction, etc.
 TYPE 3: KINASE-LINKED AND RELATED
RECEPTORS
• These membrane receptors are quite different in
structure and function from either the ligand-gated
channels or the GPCRs.
• They mediate the actions of a wide variety of protein
mediators, including growth factors and cytokines and
hormones such as insulin and leptin.
• Most of these receptors are large proteins consisting
of a single chain of up to 1000 residues, with a single
membrane-spanning helical region, associated with a
large extracellular ligand-binding domain, and an
intracellular domain of variable size and function.
 TYPES
• The tyrosine kinase receptors (TKR) have a
large extracellular agonist binding domain,one
transmembrane segment and an intracellular
domain.
• The receptors can be divided into two groups:
• (1) Those that contain the tyrosine kinase as an
integral part of the intracellular domain; and
• (2) Those that are associated with a Janus
kinase (JAK).
• Examples of the former group are
• the insulin receptor family,
• the fibroblast growth factor (FGF) receptor
family and
• Examples of the latter are the cytokine
receptor family such as
• the erythropoietin (EPO) receptor and
• the thrombopoietin (TPO) receptor.
• However, both groups share the same
mechanism of activation:
• Upon agonist binding two intracellular kinases
are brought together which will initiate
autophosphorylation of tyrosine residues of the
intracellular tyrosine kinase domain .
• This will attract other proteins (e.g.
Shc/Grb2/Sos and STAT for the two receptor
groups, respectively) which are also
phosphorylated and this will initiate protein
cascades which will ultimately lead to regulation
of transcriptional factors e.g. Elk-1, and thus
regulation of genes involved in e.g. cell
proliferation and differentiation.
 TYPE 4: NUCLEAR RECEPTORS
• Nuclear receptors are cellular proteins and are thus not
embedded in the membrane like the previously described
receptors.
• In contrast to the membrane bound receptors, they bind
small lipophilic hormones and function as ligand modulated
transcription factors.
• The nuclear receptors have been classified according to the
type of hormone they bind.
• Thereby, receptors have been divided into those which bind
steroids (glucocorticoids, progestestins, mineralocorticoid
androgens and estrogens) and steroid derivatives (vitamin
D3),
• non-steroids (thyroid hormone, retinoids, prostaglandins),
and
• The nuclear receptors consist of a ligand
binding domain and a DNA binding domain.
• Upon activation, the two receptors dimerize, as
homo- or heterodimers, and bind to specific
recognition sites on the DNA.
• Co-activators will then associate with the
dimeric receptor and initiate transcription of
the target gene(s).
• Each receptor recognize specific DNA
sequences, also known as the hormone
response elements, which are located upstream
of the genes that are regulated.