PHAR0004/9/7 - General and Systematic Pharmacology/

Intermediate Pharmacology/Introductory Pharmacology

RECEPTOR STRUCTURE Professor Neil Millar

RECEPTOR DIVERSITY

The term ‘receptor’ is used to describe a diverse group of biological molecules (including T-cell
receptors etc.) but here we will consider only proteins that are receptors for neurotransmitters,
hormones, growth factors and other inter-cellular (cell-to-cell) messengers. Such receptors are also
the site of action of many pharmacologically important drugs. Receptors can usefully be subdivided
into four structurally distinct families: 1) ligand-gated ion channels (LGICs) such as nicotinic
acetylcholine receptors, 2) G-protein- receptors (GPCRs) such as muscarinic acetylcholine
receptors, 3) kinase-linked and related receptors such as the insulin receptor and 4)
intracellular/nuclear receptors such as the oestrogen receptor. The first three of these receptor
families are transmembrane proteins and participate in transmembrane signalling events. In contrast,
the family of intracellular receptors are soluble proteins that function by regulating gene transcription
in the cell’s nucleus.

RECEPTOR NOMENCLATURE

Prior to molecular cloning, receptors were often classified on the basis of their pharmacological
properties. For example, receptors for acetylcholine were classified as either “nicotinic” or
“muscarinic” on the basis of their sensitivity to nicotine (from tobacco) or to muscarine (from the
poisonous mushroom Amanita muscaria). Receptors are often sub-classified further on the basis of
pharmacological differences (e.g. M1 and M2 type muscarinic receptors or  and  adrenoreceptors).
In some cases, subscript letters are used to distinguish receptor subtypes (e.g. GABAA and GABAB
receptors). Unfortunately, the nomenclature of receptors and of receptor subunits in the scientific
literature is not always consistent. This has been a particular problem for the nomenclature of some
LGIC subunits, although there have been attempts in recent years to standardise such nomenclature
(see, for example, Collingridge et al. 2009 Neuropharmacol. 56, 2-5).

LIGAND-GATED ION CHANNELS (LGICs)

Ligand-gated ion channels (LGICs) are multi-subunit (oligomeric) proteins containing an integral
ion-channel pore. These are sometimes referred to as “agonist-activated ion channels”,
“neurotransmitter-gated ion channels” or “ionotropic receptors”. The LGIC family includes receptors
for several neurotransmitters e.g. acetylcholine (ACh), glutamate (Glu), glycine (Gly), γ-
aminobutyric acid (GABA), 5-hydroxytryptamine (5-HT or serotonin). LGICs are frequently sub-
divided into three structurally distinct sub-families: 1) "Cys-loop" receptors, (e.g. nAChRs,
GABAARs and 5-HT3R), 2) glutamate (Glu) receptors (NMDA, AMPA and kainate GluRs) and 3)
P2X receptors (receptors for ATP). Transmembrane signalling occurs by the opening of an integral
ion channel and the flow of ions (e.g. Na+, Ca2+ or Cl-) along their electrochemical gradients (i.e. into
or out of the cell). The consequence is either a membrane depolarisation (due to the opening of
excitatory receptors, e.g. nAChRs) or membrane hyper-polarisation (opening of inhibitory receptors,
e.g. GABAARs)

LGIC structure
LGICs are oligomeric complexes in which multiple (3-5) subunits co-assemble in a ring structure to
form a central ion channel pore. Members of the Cys-loop family are pentamers, whereas glutamate
receptors are tetramers and P2X receptors and trimers. Individual LGIC subunit contains an
extracellular (agonist-binding) domain and multiple (2-4) transmembrane domains.

Department of Neuroscience, Physiology and Pharmacology, UCL

LGIC subunit diversity
There is considerable subunit diversity within most classes of LGIC. Subunits can co-assemble to
generate a variety of pharmacologically distinct receptors, either homomeric (containing only a single
type of subunit) or heteromeric (containing two or more different subunit sub-types). Subunit
composition may also be influenced by developmental changes - the subunit composition of nAChRs
expressed in foetal muscle is (α1)2β1γδ but in adult muscle is (α1)2β1δε. In some cases, subunit
diversity can be further increased by alternative splicing or by RNA editing of subunit mRNAs.

G-PROTEIN-COUPLED RECEPTORS (GPCRs)

G-protein-coupled receptors (GPCRs) constitute a very large family of receptors, each containing
seven membrane-spanning domains. GPCRs are sometimes also referred to as “seven-transmembrane
(7TM) receptors” or “metabotropic receptors”. The GPCR family includes receptors for several
neurotransmitters, including ACh (muscarinic AChRs), Glu (metabotropic GluRs) and GABA
(GABABRs) and for numerous other signalling molecules (e.g. hormones and odorant molecules).
[Note: several neurotransmitters activate both GPCRs and LGIC receptors]. As with LGICs, there are
often several receptor subtypes for each class of GPCR. For example, several different subtypes have
been identified of mAChRs (M1-M5), mGluRs (mGluR1-R7) and of G-protein-coupled 5HT
receptors (5-HT1R, 5-HT2R, 5-HT4R – 5-HT7R). [Note: 5-HT3R is a LGIC].

GPCR function
Unlike LGICs, GPCRs do not contain integral ion channels. Transmembrane signalling is effected by
the interaction of GPCRs with intracellular G-proteins. G-proteins are heterotrimeric complexes
(composed of α, β and γ subunits) that catalyse the inter-conversion of the guanine nucleotides GTP
and GDP. Activation of GPCRs leads to the release of GTP-bound α subunits that then diffuse away
and activate various enzymes and ion channels. Different GPCRs couple to a variety of different G-
proteins, which contributes to the diversity of GPCR function. Activation of G-proteins, via agonist-
activation of GPCRs, may result changes in intracellular cAMP concentration (by activation or
inhibition of adenylate cyclase), or modulation of ion channels (e.g. activation of mAChRs causing
opening of cardiac K+ channels). [More details of signalling via G-proteins will be provided in
subsequent lectures].

GPCR structure
Previously all GPCRs were thought to be composed of only one subunit, but there is evidence that at
least some GPCRs (e.g. GABAB receptors) form subunit dimers. GPCR subunits contain an
extracellular N-terminus and an intracellular C-terminus. Small ligands, such as noradrenaline
(norepinephrine), interact with amino acid residues that are located in a cleft between the α-helical
transmembrane domains. The third intracellular loop region appears to be critical for interaction of
GPCRs with intracellular G-proteins.

KINASE-LINKED AND RELATED RECEPTORS

This family of receptors, like the LGICs and GPCRs, are transmembrane proteins but contain only a
single transmembrane domain. They act as receptors for a wide range of signalling molecules (all of
which are peptides) including peptide hormones (e.g. insulin), growth factors (e.g. epidermal growth
factor; EGF) and cytokines. They contain an extracellular ligand-binding domain and an intracellular
effector domain. These receptors are often sub-divided on the basis of their signalling/catalytic
activity. Examples include tyrosine kinase receptors, tyrosine kinase-linked receptors,
serine/threonine kinase receptors and guanylate cyclase receptors. Typically, the intracellular domain
of the receptor has catalytic (e.g. tyrosine kinase) activity. Some receptors lack an intrinsic catalytic
activity but bind to and activate cytosolic kinases after receptor activation. A common mechanism of
action is for ligand binding to cause receptor dimerization. In receptors with intrinsic tyrosine kinase
activity this can result in auto-phosphorylation of tyrosine residues. This, in turn, can lead to binding
of a group of intracellular proteins (called “SH2-domain” proteins). Many SH2-domain proteins are
themselves protein kinases, or have other catalytic (e.g. phospholipase) activity. [More details of the
functional properties of these receptors will be provided in subsequent lectures].

INTRACELLULAR/NUCLEAR RECEPTORS

This family of receptors differ from the three families of receptor described previously, in that they
are soluble (cytoplasmic or intra-nuclear) proteins, rather than transmembrane proteins. Intracellular
receptors include receptors for steroid hormones and act by regulating gene transcription. Steroid
hormones are small hydrophobic molecules that are able to cross the plasma membrane and interact
with intra-cellular receptors. Receptors in this family are monomeric proteins which contain a both
a ligand- (e.g. steroid-) binding site and also a DNA-binding site. Binding of the ligand to its receptor
induces a conformational change that promotes the formation of receptor dimers. In its dimeric (ligand
bound state) the receptor is able act as a transcription factor, by binding to specific regions of genomic
DNA known as hormone-responsive elements. The consequence of the receptor binding is to promote
binding of RNA polymerase leading to gene transcription (mRNA synthesis) of specific genes (those
linked to hormone-responsive elements). Increased mRNA synthesis, in turn, leads to greater protein
synthesis (translation). Different steroid hormones exert distinct cellular responses by binding to a
variety of receptors that regulate the transcription of specific genes and results in distinct patterns of
protein synthesis.