G protein-coupled receptor

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A Mu-opioid G protein-coupled receptor with its agonist

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Figure 1. The seven transmembrane α-helix structure of a G protein-coupled receptor.

G protein-coupled receptors (GPCRs), also known as seven transmembrane receptors, 7TM receptors, heptahelical receptors, and G protein linked receptors (GPLR), are a large protein family of transmembrane receptors that sense molecules outside the cell and activate inside signal transduction pathways and, ultimately, cellular responses. G protein-coupled receptors are only found in eukaryotes, including yeast, plants, choanoflagellates, and animals. The ligands that bind and activate these receptors include light-sensitive compounds, odors, pheromones, hormones, and neurotransmitters, and vary in size from small molecules to peptides to large proteins. G protein-coupled receptors are involved in many diseases, but are also the target of around half of all modern medicinal drugs.

Classification

GPCRs can be grouped into 6 classes based on sequence homology and functional similarity:

• Class A (rhodopsin-like)
• Class B (secretin-like)
• Class C (metabotropic/pheromone)
• Class D (Fungal pheromone)
• Class E (cAMP receptors)
• Class F (Frizzled/Smoothened)

The very large rhodopsin A group has been further subdivided into 19 subgroups (A1-A19). More recently, an alternative classification system called GRAFS (Glutamate, Rhodopsin, Adhesion, Frizzled, Taste2, Secretin) has been proposed.

Physiological roles

GPCRs are involved in a wide variety of physiological processes. Some examples of their physiological roles include:

1. the visual sense: the opsins use a photoisomerization reaction to translate electromagnetic radiation into cellular signals. Rhodopsin, for example, uses the conversion of 11-cis-retinal to all-trans-retinal for this purpose.
2. the sense of smell: receptors of the olfactory epithelium bind odorants (olfactory receptors) and pheromones (vomeronasal receptors)
3. behavioral and mood regulation: receptors in the mammalian brain bind several different neurotransmitters, including serotonin, dopamine, GABA and glutamate.
4. regulation of immune system activity and inflammation: chemokine receptors bind ligands that mediate intercellular communication between cells of the immune system; receptors such as histamine receptors bind inflammatory mediators and engage target cell types in the inflammatory response
5. autonomic nervous system transmission: both the sympathetic and parasympathetic nervous systems are regulated by GPCR pathways. These systems are responsible for control of many automatic functions of the body such as blood pressure, heart rate and digestive processes.
6. cell density sensing: A novel GPCR role in regulating cell density sensing.

Figure 2. G protein-coupled receptor mechanism.

Receptor structure

GPCRs are integral membrane proteins that possess seven membrane-spanning domains or transmembrane helices (Figure 1). The extracellular parts of the receptor can be glycosylated. These extracellular loops also contain two highly conserved cysteine residues which build disulfide bonds to stabilize the receptor structure.

Early structural models for GPCRs were based on their weak analogy to bacteriorhodopsin for which a structure had been determined by both electron and X ray-based crystallography. In 2000, the first (and to date only) crystal structure of a mammalian GPCR, that of bovine rhodopsin, was solved. While the main feature, the seven transmembrane helices, is conserved, the structure differs significantly from that of bacteriorhodopsin. Some seven transmembrane helix proteins (such as channelrhodopsin) that resemble GPCRs may contain different functional groups, such as entire ion channels, within their protein.

Ligand binding and signal transduction

GPCRs include receptors for sensory signal mediators (e.g., light and olfactory stimulatory molecules); adenosine, bombesin, bradykinin, endothelin, y-aminobutyric acid (GABA), hepatocyte growth factor, melanocortins, neuropeptide Y, opioid peptides, opsins, somatostatin, tachykinins, vasoactive intestinal polypeptide family, and vasopressin; biogenic amines (e.g., dopamine, epinephrine and norepinephrine, histamine, glutamate (metabotropic effect), glucagon, acetylcholine (muscarinic effect), and serotonin); chemokines; lipid mediators of inflammation (e.g., prostaglandins and prostanoids, platelet activating factor, and leukotrienes); and peptide hormones (e.g., calcitonin, C5a anaphylatoxin, follicle stimulating hormone (FSH), gonadotropic-releasing hormone (GnRH), neurokinin, and thyrotropin releasing hormone (TRH), and oxytocin). GPCRs which act as receptors for stimuli that have yet to be identified are known as orphan receptors.

While in other types of receptors that have been studied ligands bind externally to the membrane, the ligands of GPCRs typically bind within the transmembrane domain.

The transduction of the signal through the membrane by the receptor is not completely understood. It is known that the inactive G protein is bound to the receptor in its inactive state. Once the ligand is recognized, the receptor shifts conformation and thus mechanically activates the G protein, which detaches from the receptor. The receptor can now either activate another G protein, or switch back to its inactive state. This is an overly simplistic explanation, but suffices to convey the overall set of events.

It is believed that a receptor molecule exists in a conformational equilibrium between active and inactive biophysical states. The binding of ligands to the receptor may shift the equilibrium toward the active receptor states. Three types of ligands exist: agonists are ligands which shift the equilibrium in favour of active states; inverse agonists are ligands which shift the equilibrium in favour of inactive states; and neutral antagonists are ligands which do not affect the equilibrium. It is not yet known how exactly the active and inactive states differ from each other.

If a receptor in an active state encounters a G protein, it may activate it (Figure 2, blue protein in part B). Some evidence suggests that receptors and G proteins are actually pre-coupled. For example, binding of G proteins to receptors affects the receptor's affinity for ligands. Activated G proteins are bound to GTP.

The enzyme adenylate cyclase (Figure 2, green protein in panel C) is an example of a cellular protein that can be regulated by a G protein. Adenylate cyclase activity is activated when it binds to a subunit of the activated G protein (Figure 2, Panel D). Activation of adenylate cyclase ends when the G protein returns to the GDP-bound state (Figure 2, panels E and A).

GPCR signaling without G proteins

In the late 1990s, evidence began accumulating that some GPCRs are able to signal without G proteins. The ERK2 mitogen-activated protein kinase, a key signal transduction mediator downstream of receptor activation in many pathways, has been shown to be activated in response to cAMP-mediated receptor activation in the slime mold D. discoideum despite the absence of the associated G protein α- and β-subunits.

In mammalian cells the well-studied β2-adrenoceptor has been demonstrated to activate the ERK2 pathway after arrestin-mediated uncoupling of G protein mediated signalling. It therefore seems likely that some mechanisms previously believed to be purely related to receptor desensitisation are actually examples of receptors switching their signalling pathway rather than simply being switched off.

In kidney cells, the bradykinin B2 receptor has been shown to interact directly with a protein tyrosine phosphatase. The presence of a tyrosine-phosphorylated ITIM (immunoreceptor tyrosine-based inhibitory motif) sequence in the B2 receptor is necessary to mediate this interaction and subsequently the antiproliferative effect of bradykinin.

Receptor regulation

GPCRs become desensitized when exposed to their ligand for a prolongued period of time. There are two recognized forms of desensitization: 1) homologous desensitization, in which the activated GPCR is downregulated and 2) heterologous desensitization, where the activated GPCR causes downregulation of a different GPCR. The key reaction of this downregulation is the phosphorylation of the intracellular (or cytoplasmic) receptor domain by protein kinases.

Phosphorylation by cAMP-dependent protein kinases

Cyclic AMP-dependent protein kinases (protein kinase A) are activated by the signal chain coming from the G protein (that was activated by the receptor) via adenylate cyclase and cyclic AMP (cAMP). In a feedback mechanism, these activated kinases phosphorylate the receptor. The longer the receptor remains active, the more kinases are activated, the more receptors are phosphorylated.

Phosphorylation by GRKs

The G protein-coupled receptor kinases (GRKs) are protein kinases that phosphorylate only active GPCRs.

Phosphorylation of the receptor can have two consequences:

1. Translocation. The receptor is, along with the part of the membrane it is embedded in, brought to the inside of the cell, where it is dephosphorylated and then brought back. This mechanism is used to regulate long-term exposure, for example, to a hormone.
2. Arrestin linking. The phosphorylated receptor can be linked to arrestin molecules that prevent it from binding (and activating) G proteins, effectively switching it off for a short period of time. This mechanism is used, for example, with rhodopsin in retina cells to compensate for exposure to bright light. In many cases, arrestin binding to the receptor is a prerequisite for translocation.

Receptor oligomerization

It is generally accepted that G protein-coupled receptors can form homo- and/or heterodimers and possibly more complex oligomeric structures, and indeed heterodimerization has been shown to be essential for the function of receptors such as the metabotropic GABA(B) receptors. However, it is presently unproven that true heterodimers exist. Present biochemical and physical techniques lack the resolution to differentiate between distinct homodimers assembled into an oligomer or true 1:1 heterodimers. It is also unclear what the functional significance of oligomerization might be, although it is thought that the phenomenon may contribute to the pharmacological heterogeneity of GPCRs in a manner not previously anticipated. This is an actively studied area in GPCR research.

Plants

GCR2 is a G protein-coupled receptor for the plant hormone abscissic acid that has been identified in Arabidopsis thaliana. Another putative receptor is GCR1 for which no ligand has been identified yet. A novel GPCR containing a lipid kinase domain has recently been identified in Dictyostelium that regulates cell density sensing.

See also

• Orphan receptor

External links

• A phylogenetic tree of all human GPCRs showing family relationships from Vassilatis, et al.

• IUPHAR GPCR Database
• G Protein-Coupled Receptor Database (GPCRDB)
• G-protein-coupled+receptors at the U.S. National Library of Medicine Medical Subject Headings (MeSH)
• Wikipedia:MeSH D12.776#MeSH D12.776.543.750.100 --- receptors.2C g-protein-coupled
• UMich Orientation of Proteins in Membranes families/superfamily-6