SIGNALING THROUGH

ENZYME-COUPLED
CELL-SURFACE RECEPTORS
Introduction
• Enzyme-coupled receptors are single transmembrane domain
containing proteins with their ligand-binding domain on the outer
surface of the plasma membrane.
• Their cytosolic domain either has intrinsic enzyme activity or
associates directly with an enzyme.
• GPCRs and enzyme-coupled receptors often activate some of the
same signaling pathways, and there is usually no obvious reason
why a particular extracellular signal utilizes one class of receptors
rather than the other.
Classes of enzyme-coupled receptors
There are six principal classes of enzyme-coupled receptors

• Directly phosphorylate specific tyrosines on

1. Receptor themselves and on a small set of intracellular
tyrosine kinases signaling proteins.

2. Tyrosine- • Have no intrinsic enzyme activity but directly

kinase-associated recruit cytoplasmic tyrosine kinases to relay the
signal.
receptors

• Directly phosphorylate specific serines or

3. Receptor
threonines on themselves and on latent gene
serine/threonine regulatory proteins with which they are
kinases associated.
 4. Histidine- • Activate a two-component signaling pathway in which
the kinase phosphorylates itself on histidine and then
kinase-associated immediately transfers the phosphoryl group to a
receptors second intracellular signaling protein.

• Directly catalyze the production of cyclic GMP in the

5. Receptor cytosol, which acts as a small intracellular mediator in
guanylyl cyclases much the same way as cyclic AMP.

• Remove phosphate groups from tyrosines of specific

6. Receptor like intracellular signaling proteins. (They are called
tyrosine “receptorlike” because their presumptive ligands have
not yet been identified, and so their receptor function
phosphatases is unproven).
1. Receptor tyrosine kinases
Introduction
• Receptor tyrosine kinases (RTKs) are the most numerous class of the enzyme-coupled receptors.
• Many extracellular signal proteins that act through RTKs, include
• Epidermal growth factor (EGF),
• Platelet-derived growth factor (PDGF),
• Fibroblast growth factors (FGFs),
• Hepatocyte growth factor (HGF),
• Insulin,
• Insulinlike growth factor-1 (IGF1),
• Vascular endothelial growth factor (VEGF),
• Macrophage-colony-stimulating factor (MCSF), and
• Nerve growth factor (NGF).
• Additionally many cell-surface-bound extracellular signal proteins also act through RTKs. For
example Ephrins, the largest class of such membrane-bound ligands, with eight identified in
humans that act through Eph receptors, the most numerous RTKs, with thirteen genes encoding
them in humans.
 Structural subfamilies of RTKs
• There are about 60 genes encoding human RTKs. These receptors can
be classified into more than 16 structural subfamilies,

Some subfamilies of RTKs

that operate in mammals,
Table showing some of the RTKs ligands and their resultant responses
RTKs activation
• In all cases, the binding of the
signal protein to the ligand-
binding domain of the receptor,
on the outside of the cell, causes
the receptor chains to dimerize.
• This bring the kinase domains of
two receptor chains together (an
example of induced proximity), so
that they can become activated
and cross-phosphorylate each
other on multiple selected
tyrosine side chains, a process
referred to as
transautophosphorylation.
Phosphotyrosine docking sites

Phosphorylated Tyrosines on RTKs serve as Docking Sites for

Intracellular Signaling Proteins. The cross-phosphorylation of adjacent
cytosolic tails of RTKs contributes to the receptor activation process in
two ways.
1. Phosphorylation of tyrosines within the kinase domain increases the kinase
activity of the enzyme.
2. phosphorylation of tyrosines outside the kinase domain creates high-
affinity docking sites for the binding of specific intracellular signaling
proteins that contains a specific phosphotyrosine-binding domain.
• Once bound to the activated RTK, a
signaling protein may itself become
phosphorylated on tyrosines and
thereby be activated.
• In many cases, however, the binding
alone may be sufficient to activate the
docked signaling protein, by either
inducing a conformational change in the
protein or simply bringing it near the
protein that is next in the signaling
pathway.
• Thus, transautophosphorylation triggers
the transient assembly of an intracellular
signaling complex, which can then relay The docking of intracellular signaling
the signal onward, often along multiple proteins on phosphotyrosines on an
activated RTK
routes, to various destinations in the cell
 Because different RTKs
bind different combinations of these
signaling proteins, they activate
different responses
Docking proteins

• Most of the phosphotyrosine docking sites generated by ligand

binding are not on the receptor itself but on a specialized docking
protein for example insulin receptor substrate-1 (IRS1).
• The activated receptor first transautophosphorylate its kinase
domains, which then phosphorylate docking protein on multiple
tyrosines, thereby creating many more docking sites than could be
accommodated on the receptor alone.
• Some other RTKs use docking proteins in a similar way to enlarge the
size of the signaling complex.
• How the intracellular signaling proteins binds to the phosphotyrosine
residues either on the receptor or on the docking protein?

• How the specific intracellular signaling proteins identify the specific

phosphorylated residue ?
 Phosphotyrosine-Dependent Protein–
Protein Interactions
• The intracellular signaling proteins can bind to the phosphotyrosines either
on activated RTKs or on docking proteins such as IRS1, and help to relay the
signal onward, mainly through chains of protein–protein interactions.
• Some of these docked proteins are enzymes like
• phospholipase C-Ƴ (PLCƳ), which functions in the same way as phospholipase C-β—
activating the inositol phospholipid signaling pathway, increase cytosolic Ca 2+ levels
and activate PKC.
• Cytoplasmic tyrosine kinase, Src, which phosphorylates other signaling proteins on
tyrosines.
• Phosphoinositide 3-kinase (PI 3-kinase), which phosphorylates mainly lipids, the
phosphorylated lipids then serve as docking sites to attract various signaling proteins
to the plasma membrane.
Interaction domains: Phosphotyrosine-binding domains
• Signaling proteins that bind to phosphotyrosines, either on activated
RTKs or on proteins docked on them, although have varied structures
and functions but usually share highly conserved phosphotyrosine-
binding domains.
• Two of these domains have been identified thus far
• SH2 domains (for Src homology region) or,
• PTB domains (for phosphotyrosine-binding) less common.
• By recognizing specific phosphorylated tyrosines, these small
interaction domains enable the proteins that contain them to bind to
activated RTKs, as well as to many other intracellular signaling
proteins that have been transiently phosphorylated on tyrosines.
 SH2 Domain
• SH2 domains are composed of approximately
100 amino acids and contain a conserved
binding-pocket that accommodates a
phosphorylated tyrosine residue. More than
110 SH2 domains are encoded by the human
genome. They mediate a large number of
phosphorylation-dependent protein–protein
interactions.
• These interactions occur following
phosphorylation of specific tyrosine residues.
The specificity of the interactions is
determined by the amino acid sequence
immediately adjacent to the phosphorylated
tyrosine residues. For example, the SH2
domain of the Src protein-tyrosine kinase
recognizes pTyr-Glu-Glu-Ile, whereas the SH2
domains of PI 3-kinase bind to pTyr-Met-X-Met
(in which X can be any residue)
Interaction domains: SH3 and PH

• In addition to SH2 and PTB domains that bind to phosphorylated

tyrosines in a particular peptide sequence on activated receptors or
intracellular signaling proteins, there are other interaction domains,
for example
• Src homology 3 (SH3) domain: bind to short proline-rich amino acid
sequences
• Pleckstrin homology (PH) domain: bind to the charged head groups of specific
phosphoinositides that are produced in the plasma membrane in response to
an extracellular signal.
 The binding of SH2-containing intracellular signaling proteins to an activated
PDGF receptor.

many signaling proteins also contain

other interaction domains that allow
them to interact specifically with other
proteins as part of the signaling
process. For example SH3 domain.

binds to proline-
rich motifs in
intracellular
proteins
These numbers
indicate the positions
of the tyrosines in
the polypeptide
chain

Phosphotyrosine docking sites. Mutation

of specific tyrosine will prevents the
binding and activation of particular
signaling protein.
Adaptors
• Some signaling proteins consist solely of two or more interaction
domains and function only as adaptors.
• Adaptor proteins function as linkers that enable two or more signaling
proteins to become joined together as part of a signaling complex.
• Adaptor proteins contain an SH2 domain and one or more additional
protein–protein interaction domains. For instance, the adaptor
protein Grb2 contains one SH2 and two SH3