Cell signaling receptor

All cells in a multicellular organism are constantly exposed to a variety of extracellular

signals that they need to interpret and translate into an appropriate response to their
environment. These signals can be soluble factors generated locally (for example,
synaptic transmission) or distantly (for example, hormones and growth factors), ligands
on the surface of other cells, or the extracellular matrix itself. To achieve this, cells
maintain a diversity of receptors on their surface that respond specifically to individual
stimuli. These receptors fall into families, based primarily on the way in which they
generate the intracellular signals that give rise to the particular functional responses.
Moreover, the activity of a given receptor can be modulated by other signaling pathways
in a variety of ways, generating the flexibility required of such a complex system. This
review aims to describe the function of the major classes of receptor, including G protein
coupled receptors, receptor tyrosine kinases, ligand gated ion channels, integrins, and
cytokine receptors, and to demonstrate the “crosstalk” that exists between these systems.

The ability of cells to interact with and adapt to their environment is one of the
fundamental processes of cell biology. This responsiveness is achieved
predominantly through the expression at the cell surface of a repertoire of specific
receptors that are sensitive to the composition of the surrounding environment. The
signals that trigger these receptors can be presented in a variety of contexts—for
example, soluble factors (chemicals, polypeptides, proteins, sugars, etc), a ligand
bound to another cell, or the extracellular matrix itself. The receptors then
transduce these extracellular signals across the plasma membrane and, through the
activation of intracellular signalling pathways, bring about the appropriate
functional response. Moreover, cells have developed sophisticated systems to
integrate inputs from multiple signals.

In this review, we will discuss the various means by which different classes of cell
surface receptors are triggered, the mechanisms by which the signals are
terminated, and the ways in which the activities of the receptors can be modulated.
We begin by looking at the diverse family of cytokine receptors that play a key role
in regulating the function of the haematopoietic system and in coordinating immune
responses.
Cytokine receptor:-

on numerous cell types and can interact with, induce, or inhibit other cytokines, leading
to the development of the concept of a cytokine network controlling those biological
processes that involve cellular interactions.1 Cytokines can regulate diverse biological
processes such as cell death and survival, cell proliferation, and cellular differentiation 2;
moreover, different cytokines can act on the same cell type to induce a similar response.
These abilities are dependent in part upon the multimeric nature of the cell receptors for
cytokines,3 but also illustrate a fundamental issue in receptor mediated signalling, the
means by which diverse extracellular stimuli are integrated into a coherent response.
RECEPTOR CLASSIFICATION

The molecular revolution in biology has allowed the cloning of numerous cytokines and
their cognate receptors, and it has become clear that the receptors can be classified into
families according to protein sequence homologies in their extracellular domains. 4

One of the largest families comprises the type I or haematopoietin receptors, which
respond to cytokines such as interleukin 2 (IL-2), IL-4, IL-5, and IL-6, prolactin growth
hormone, and granulocyte macrophage colony stimulating factor (GM-CSF). These
receptors share features in two domains located in the extracellular portion. This
homologous region consists of two globular structures that fold into a barrel-like
structure, with a hinge region between them, which functions as a ligand binding
pocket. All type 1 receptors contain a single membrane spanning domain and a
cytoplasmic domain. The characterisation of the composition of type I receptors has led
to a better understanding of the functional redundancy attributed to their cytokine ligands.
High affinity receptors are made up of two chains, namely α and β. The α chain is the
cytokine specific subunit, the β chain alone cannot recognise a particular cytokine.
However, it forms the high affinity receptor upon binding to the α chain, and is then
responsible for transducing the signal. Thus, the fact that some receptors share the same β
chain can explain some of the coexistent biological activities of individual cytokines. The
multimeric composition of this family of receptors can therefore allow the classification
of cytokines based on the presence of a common signal transducing element—for
example, the gp140 subgroup which comprises the receptors for IL-3, GM-CSF, and IL-
5. However, it should be noted that functional redundancy is also achieved by the use of
common downstream signalling molecules, as will be described for the receptors for
tumour necrosis factor (TNF) and IL-1.
Class II cytokine receptors show some structural similarities to type I, but differ in the
presence of characteristic cysteine pairs located in the extracellular domain. A key
member of this family is the receptor for interferon.

Class III cytokine receptors comprise the proteins related to the TNF receptor. This large
and important family is characterised primarily by the presence of multiple cysteine rich
extracellular domains, varying from two to six repeats. However, the family can be
subdivided into two groups: those without appreciable homology in the cytoplasmic
domains (such as the receptors for nerve growth factor (NGF) and CD40) and those that
contain an 80 amino acid cytoplasmic “death domain”. So far, only one member of the
family, namely CD120a, has been subjected to crystallographic analysis, the results of
which support the hypothesis that a trimeric ligand complex binds to three receptor
molecules.

Many of the members of this family exist in soluble forms; indeed osteoprotegrin, the
focus of much recent interest (as a result of its role in bone formation), exists only as a
soluble molecule. These soluble receptors consist of the extracellular portion of the
corresponding full length receptor, and seem to function by competing with the
membrane bound receptor for soluble ligand. Interestingly, some viruses have exploited
this inhibitory potential as a means of defence against the antiviral activities of TNF
ligands. For example, poxviruses produce soluble forms of CD120b and CD30.

Extracellular matrix :-

Thus far, we have focused on the regulation of crosstalk between cells and the key role
played by cytokine receptors; however, cell survival and proliferation require contact
with the extracellular matrix, an interaction that is regulated primarily by another family
of cell surface receptors, the integrins. The extracellular matrix exerts profound control
over cells; integrins function to attach cells to the matrix and mediate mechanical and
chemical signals. Growth factor receptors and ion channels are regulated by these signals,
which converge on diverse functions such as apoptosis, proliferation, and differentiation.

Integrin receptors are composed of two subunits, namely α and β; each αβ combination
has its own binding specificity and signalling properties. Most recognise several
extracellular matrix proteins, and conversely several matrix proteins such as fibronectin
and collagens bind to several integrins. Integrin extracellular binding activity is regulated
from the inside of the cell, whereas binding of the extracellular matrix induces signals
that are transmitted into the cell.
The cytoplasmic portions of integrins are generally short and are without intrinsic
enzymatic activity. Consequently, the signals are transduced by associating with adaptor
proteins such as cytoplasmic kinases and transmembrane growth factor receptors.
Various kinase families can be activated, including tyrosine kinases, such as FAK and
Fyn, and src family kinases. As the integrins bind to the extracellular matrix they cluster
in the plane of the cell membrane, an event that results in the assembly of actin filaments.
This reorganisation of actin filaments into larger stress fibres in turn causes more integrin
clustering, creating a positive feedback system. Consequently, extracellular matrix
proteins, integrins, and cytoskeletal proteins assemble into aggregates on either side of
the membrane, which can be visualised by confocal microscopy.

The cooperation between integrins and growth factor receptors is necessary for optimal
activation. For example, the receptor for insulin undergoes maximal activation only under
appropriate cell attachment conditions. It also appears that certain integrins can
preferentially associate with particular growth factor receptors. Growth factor receptors
themselves constitute a large and important family and illustrate several interesting
additional principles.

Tyrosine kinase :-

Polypeptide growth factors such as epidermal growth factor (EGF), and platelet derived
growth factor (PDGF) exert profound effects on cellular proliferation, maturation, and
function, particularly during development and in the response to injury in the adult.
Malfunction or dysregulation of these powerful systems contributes to the development
of many neoplastic growths, where they provide a persistent mitogenic signal for the
transformed cells. The growth factor receptors form a family that has intrinsic protein
tyrosine kinase activity in the cytoplasmic domains. The receptors all have a single
membrane spanning helix and an extracellular ligand binding domain of diverse
structure. When a growth factor binds to its receptor, it induces both receptor
dimerisation and an increase in the activity of the kinase. The intracellular region of the
activated receptor then becomes extensively autophosphorylated, and these
phosphorylated tyrosines form specific docking sites for intracellular molecules that
contain SH2 domains. In this way, a large signalling complex is assembled rapidly
around activated receptors, from which a variety of signal transduction cascades emanate.
For instance, in the activated PDGF β receptor, two phosphotyrosine residues close to the
membrane mediate binding of Src family kinases and STAT (signal transducer and
activator of transcription) molecules, two sites at the C-terminus recognise phospholipase
Cγ and the tyrosine phosphatase SHP-2, and sites in the middle of the cytoplasmic
domain recruit phosphatidyl inositol 3-kinase, the GTPase activating protein of ras, and
adaptors such as Grb2, Nck, and Shc. Many of these proteins are substrates for the
receptor kinase and become phosphorylated. The multiplicity of components rapidly
activated in this way underpins the profound and diverse effects growth factors such as
PDGF have on their target cells because each signalling pathway that is activated can
initiate or amplify one or more functional responses. Moreover, many growth factor
ligands induce heterodimerisation of multiple receptors, and the signalling pathways and
functional responses subsequently activated change with receptor composition, providing
a mechanism to generate a large number of different cellular responses from a limited
number of receptors.

Concomitant with the initiation of signal transduction, activated receptors are internalised
rapidly by the endocytic pathway and subsequently degraded in the lysosomes. In
addition, receptor phosphorylation appears to acts as a signal for ubiquitinylation, thus
promoting degradation of receptors by a proteasome dependent pathway. Consequently,
ligand binding causes a downregulation of surface receptors and therefore an attenuation
of the signal transduction cascades, thus limiting the degree of cellular activation.

Ligand gated ion channel:-

The maintenance of the cell membrane as a barrier to the flux of ions plays a central role
in cellular physiology, particularly in the nervous system. Therefore, it is not surprising
that many receptors function by altering the permeability of membranes to specific ions.
Certain receptors in the central nervous system, such as the GABA A, 5HT3, glycine, and
nicotinic acetylcholine (nACh) receptors all function in a similar fashion to ligand gated
ion channels. These receptors are pentameric complexes, assembled from several similar
(in some cases identical) subunits, each of which has a large extracellular (synaptic)
region, several membrane spanning regions, and small intracellular domains. A single
transmembrane α-helix from each subunit is used to form a central pore through which
the ions pass. The glycine receptor has an additional cytoplasmic component, gephyrin,
which links the receptor to the cytoskeleton and other signalling components. Ligand
binding occurs on the synaptic domain of one or more of the subunits, and induces a
conformational change in the receptor from a “closed” state, which is impermeable to
ions, into an “open” state, allowing ions to flow through the central pore. This is followed
by a rapid transition to a third state, referred to as “desensitised”, where the channel is
closed and the receptor unresponsive to further stimulation. This desensitised channel
then slowly resets into the closed state ready for reactivation.
By virtue of the size and charge of residues lining the pore, different receptors show
selective permeability for different ions. The glycine and GABA A receptors are anion
channels that, when open, allow the movement of chloride ions. In contrast, the nACh
receptor is selectively permeable to monovalent cations such as sodium. The flow of ions
can initiate neuron depolarisation and hence activation (for example, the nACh receptor),
or can decrease the responsiveness of the synapse (for example, the glycine receptor).

The activity of ligand gated ion channels can be altered in several ways. Allosteric
modulators (such as zinc ions for the glycine receptor) can increase or decrease the
apparent affinity of the receptors for their cognate ligands More interestingly, the
cytoplasmic portion of the receptors can be phosphorylated by a variety of kinases
including protein kinase A (PKA), PKC, calmodulin dependent kinase II (CAMKII) and
Src. The functional consequences of this phosphorylation are diverse. Serine
phosphorylation of the nACh receptor by PKA appears to increase the rate of receptor
desensitisation. In contrast, tyrosine phosphorylation of the glutamate receptor enhances
the functioning of the channel. Thus, prior activation of intracellular signalling pathways
in response to other stimuli can modulate the sensitivity of neurons to ligands such as
glycine or ACh. This complexity might be the key to synaptic plasticity, and serves to
illustrate the way in which cells use signal transduction pathways to integrate the set of
signals that they receive from their environment into an appropriate coordinated cellular
response.

G – protein coupled receptor :-

The G protein coupled seven transmembrane proteins (GPCs) form one of the largest
families of cell surface receptors. They share a consistent general topology, with an N-
terminal extracellular domain, seven membrane spanning helices separated by loop
regions of varying sizes, and an intracellular C-terminal domain. GPC receptors activate
signal transduction through the coupling of their cytoplasmic domains to a family of
heterotrimeric GTP binding proteins (G proteins). Ligand binding promotes the exchange
of G protein bound GDP for GTP, and this activated G protein leaves the receptor
complex and initiates signal transduction. The GTP is hydrolysed to GDP by the intrinsic
GTPase activity of the G protein itself, providing a convenient self limiting mechanism.
However, each receptor can activate a large number of G proteins before the signal is
terminated by receptor internalisation, ensuring substantial amplification of the signal
before termination. Different GPC receptors use the same basic mechanism to act on a
wide variety of signal transduction pathways, including adenyl cyclase, tyrosine kinase
cascades, and phospholipases.
The ligands for these receptors are diverse and they mediate a large number of cell
responses as varied as retinal stimulation by light, the control of heart rate, and cellular
migration. To illustrate this diversity we will briefly examine two emerging areas of
general interest; namely, mechanisms of odorant and chemosensory receptors and
chemokine receptors.

Chemosensation in higher organisms consists of taste and smell. In mammals, olfactory

sensory neurons are located in the nasal cavity. Odorants activate specific receptors on
the cilia of these neurons. These molecular receptors are invariably G protein coupled
seven transmembrane proteins. In the mouse and rat, the repertoire of odorant receptors is
estimated to be about 1% of the genome, more than the T cell receptor and
immunoglobulin families combined. These genes were first identified by exploiting
several features and assumptions about their likely structure, in particular the
conservation of certain amino acid motifs within the seven transmembrane region.
However, compared with the other gene families already discussed, information about
these genes is still fragmentary. Recent genetic manipulation experiments have suggested
that, in vertebrates, odorant receptors might play a dual role as both chemosensors and as
molecular addressers that guide axons of olfactory sensory neurons to their precise target
in the brain.

Chemokines are small proteins that function as chemoattractants for haematopoietic cells
and are particularly involved in the coordination of immune and inflammatory
responses. There are two principal classes of chemokines, defined by the spacing of
cysteine residues as CC and CXC chemokines. The CC chemokines include RANTES,
eotaxin, monocyte chemoattractant protein 1 (MCP-1), and macrophage inflammatory
protein 1α (MIP-1α), whereas the CXC chemokines are exemplified by IL-8 and Gro-α.
There is an increasing number of chemokine receptors, all of which appear to be
particularly promiscuous, with each receptor binding to multiple ligands and most of the
chemokines using a variety of receptors. Thus, RANTES will activate CCR1, CCR3, and
CCR5, whereas CCR1 will bind RANTES, MCP-1 and MIP-1α. However, there is no
crossreactivity between the CC and CXC families.

The chemokine receptors are classic GPC receptors, functioning through heterotrimeric G
proteins. Activated chemokine receptors are internalised rapidly by an endosomal
pathway, leading to desensitisation. Moreover, unlike many cytokine receptors, the
repertoire of chemokine receptor expression at the surface of any one cell type is
particularly variable, being sensitive to many kinds of cellular activation. Thus, the
responsiveness of cells to chemokines is very dependent on the environment to which
they have been exposed. This is exactly what one would expect, given their role as part of
the complex coordination of immune responses, and illustrates another mechanism that
cells use to adapt their behaviour according to multiple inputs.