Cell surface receptors

Classes of cell surface receptors

Hydrophilic hormones such as peptides, catecholamines cannot
penetrate the target cell’s plasma membrane, so they bind to receptor
on cell’s surface and alter them indirectly.
Most classes of cell surface receptors belong to one of the 3 classes.
1. G-Protein coupled Receptors
2. Enzyme-coupled Receptors
3. Ion channel coupled Receptors
Introduction
• G protein-coupled receptors (GPCRs) are so named because they
interact with G proteins.
• Members of the GPCR superfamily are also referred to as seven-
transmembrane (7TM) receptors because they contain seven
transmembrane helices
Topology of GPCR
• Their amino-terminus is present on the outside of the cell
• The seven helices that traverse the plasma membrane are connected
by loops of varying length
• The carboxyl-terminus is present on the inside of the cell (Figure
15.4).
• There are three loops present on the outside of the cell that, together,
form the ligand-binding site.
• There are also three loops present on the cytoplasmic side of the
plasma membrane that provide binding sites for intracellular signaling
proteins. G proteins bind to the third intracellular loop.
Ligands of GPCRs
• GPCRs constitute the single largest superfamily of proteins encoded by
animal genomes.
• Included among the natural ligands that bind to GPCRs are a diverse array
of
• Hormones (both plant and animal),
• Neurotransmitters,
• Opium derivatives,
• Chemoattractants (e.g., molecules that attract phagocytic cells of the immune
system),
• Odorants and tastants (molecules detected by olfactory and gustatory receptors
eliciting the senses of smell and taste,
• Photons
G Protein
• These proteins are referred to as G proteins because they bind
guanine nucleotides, either GDP or GTP.
• They are described as heterotrimeric because all of them consist of
three different polypeptide subunits, called, α, β, and Ƴ (Figure 15.4).
• This property distinguishes them from small, monomeric G proteins,
such as Ras.
• Heterotrimeric G proteins are held at the plasma membrane by lipid
chains that are covalently attached to the α and Ƴ subunits.
• The guanine nucleotide-binding site is present on the Gα subunit.
Types of G protein
• Based upon the Gα subunit and the effectors to which they couple,
heterotrimeric G proteins have been classified into four families based
on the amino acid sequence relatedness of the α subunits
General overview of Signal Transduction
by GPCRs

Step 1: The ligand

binds to the receptor,
altering its
conformation and
increasing its affinity
for the G protein to
which it binds.
Step 2: The interaction with the receptor induces a
conformational change in the α subunit of a G protein,
causing the release of GDP, which is followed by binding
of GTP. In its GTP-bound conformation, the G α has a low
affinity for GβƳ, leading to its dissociation from the
complex.
Step 3: The Gα subunit dissociates from the GβƳ complex
and binds to an effector (e.g phospholipase C, cyclic
GMP phosphodiesterase, adenylyl cyclase), activating
the effector.
Step 4: Activation of the effector (adenylyl cyclase in this
case) leads to the production of the second messenger
cAMP.
Step 5: The α subunit has GTPase
activity (hydrolyzing GTP to GDP and
inorganic phosphate, Pi), so once GTP is
hydrolyzed, it becomes inactive. This
results in a conformational change
causing a decrease in the Gα affinity for
the effector and an increase in the
affinity for the βƳ subunit. Thus,
following hydrolysis of GTP, the Gα
subunit will dissociate from the effector
and reassociate with the βƳ complex to
reform the inactive heterotrimeric G
protein.
• The GTPase activity is greatly
enhanced by the binding of the
Gα subunit to either the effector
protein or a specific regulator of
G protein signaling (RGS).
• RGS proteins act as a-subunit-
specific GTPase-activating
proteins (GAPs) and they help
shut off G protein-mediated
responses in all eucaryotes. There
are about 25 RGS proteins
encoded in the human genome,
each of which interacts with a
particular set of G proteins
Intracellular signaling pathways activated
by GPCRs
GPCRs activate various intracellular signaling pathways, including some that are also activated by
enzyme-coupled receptors.

1. Some G Proteins Regulate the Production of

Cyclic AMP (cAMP)

2. Some G Proteins Activate An Inositol

Phospholipid Signaling
Pathway by Activating Phospholipase C-b (PLC-β)

3. Some G Proteins Directly Regulate Ion Channels

1. cAMP Signaling pathway
• Cyclic AMP (cAMP) acts as a small
intracellular mediator in all prokaryotic and
animal cells.
• Its normal concentration in the cytosol is
about 10–7 M, but an extracellular signal can
increase this concentration more than
twentyfold in seconds.
• cAMP is synthesized from ATP by a plasma-
membrane-bound enzyme adenylyl
cyclase, and it is rapidly and continuously
destroyed by cyclic AMP
phosphodiesterases that hydrolyze cAMP
to adenosine 5’-monophosphate (5’-AMP).
• Adenylyl cyclase is a large multipass transmembrane protein with its catalytic
domain on the cytosolic side of the plasma membrane. There are at least eight
isoforms in mammals, most of which are regulated by both G proteins and
Ca2+.
• GPCRs that act by increasing cAMP are coupled to a Gs, which activates
adenylyl cyclase and thereby increases cAMP concentration.
• Response generated by increase in cAMP concentration is dependent on cell
type, with different cell types responding differently to it. However one cell
type often responds in the same way to such an increase, regardless of the
extracellular signal that causes it.
• Examples of some of the responses generated by cAMP are given in table on
next slide
Intracellular effects of cAMP
• In most animal cells, cyclic AMP exerts its effects
mainly by activating cyclic-AMP-dependent
protein kinase (PKA).
• PKA phosphorylates specific serines or
threonines on selected target proteins, including
intracellular signaling proteins and effector
proteins, thereby regulating their activity. The
target proteins differ from one cell type to
another, which explains why the effects of cAMP
vary so markedly depending on the cell type.
• In the inactive state, PKA consists of a complex
of two catalytic subunits and two regulatory
subunits.
• The binding of cAMP to the regulatory subunits
alters their conformation, causing them to
dissociate from the complex. The released
catalytic subunits are thereby activated to
phosphorylate specific target proteins
1. The released catalytic subunits of PKA can
then enter the nucleus, where they
phosphorylate the gene regulatory protein,
cAMP response element binding protein
(CREB) on a single serine.
2. Once phosphorylated, CREB recruits the
transcriptional coactivator called CREB
binding protein (CBP)
3. It then stimulates gene transcription by
binding to the cAMP response element (CRE)
in DNA .
4. This signaling pathway controls many
processes in cells, ranging from hormone
synthesis in endocrine cells to the production
of proteins required for the induction of long-
term memory in the brain.