Signal Transduction by G Protein-Coupled Receptors

Introduction Many vital physiological processes rely on the signal transduction by G protein-coupled receptors (Table 1). Signal transduction is the process by which a ligand (an extracellular signal molecule) interacts with a cellular receptor and induces a change in cellular physiology, usually by an effector (a molecule that brings about a cellular response to a signal) 1. This description explains the background knowledge on G protein-coupled receptors and G proteins, and the mechanism of receptor-mediated activation or inhibition of effectors by means of G proteins. Table 1. Physiological processes governed by G protein-coupled receptors 1. Ligand Type of G proteinEffector Physiological coupled receptors Response Epinephrine β-Adrenergic receptor Adenylyl cyclase Glycogen breakdown Serotonin Serotonin receptor Adenylyl cylase Behavioral sensitization Light Rhodopsin cGMP Visual excitation phsphodiesterase IgE-antigen Mast cell IgE receptor Phospholipase C Secretion complexes f-Met Peptide Chemotactic receptor Phospholipase C Chemotaxis Acetylcholine Muscarinic receptor Potassium channel Slowing of pacemaker activity G Protein-Coupled Receptors G protein-coupled receptors (GPCRs) are a vast family of plasma membrane receptors that contain seven transmembrane α helices (Figure 1) 1. Hundreds of different GPCRs have been discovered in eukaryotes ranging from yeast to plants and mammals. GPCRs are so named because they interact with G proteins. In fact, GPCRs translate the binding of ligands into the activation of G proteins, which ultimately induces respective physiological responses. Topology of GPCRs GPCRs usually have the following topology. Their amino (NH2) terminus is located on the outside of the cell, the seven α helices that pass through the plasma membrane are attached by polypeptide loops of varying length, and the carboxyl (CO2H) terminus is present on the inside of the cell (Figure 1) 1. These three distinct regions are often referred to as the extracellular, transmembrane, and cytosolic domains, respectively. In the extracellular domain, there exist three loops, which form a ligand-binding site. Similarly, there are also three loops present in the cytosolic domain, forming a G protein binding site. Both binding sites are critical in the signal transduction by GPCRs.

Extracellular Domain

Transmembrane Domain

Cytosolic Domain

Figure 1. A diagram showing the G protein-coupled receptor (left), the G protein (middle), and the effector (right) 1. G Proteins G proteins are also referred as guanine nucleotide-binding proteins because they bind guanine nucleotides, either as GDP or GTP. Although there are two classes of G proteins (monomers and heterotrimers), only heterotrimers are associated with GPCRs. G proteins are heterotrimers because they consist of three different polypeptide subunits: Gα, Gβ, and Gγ 2. G proteins bind to GPCRs using the Gα subunit at the G protein binding site (Figure 1). In addition, there are two states of G proteins: active and inactive. The active state consists of Gα subunit binding to GTP and separated from Gβ and Gγ subunits (often stay together and are referred to as Gβγ subunits). The inactive state, on the other hand, consists of Gα subunit binding to GDP and attached to Gβ and Gγ subunits. Because the Gα subunit contains a GTPase (an enzyme that hydrolyzes GTP into GDP) endogenously, the Gα subunit is capable of inactivating itself. The Mechanism of GPCRs The process of receptor-mediated activation or inhibition of effectors by means of G proteins is explained in eight steps below (Figure 2):

2. The binding of inactive G protein to the receptor results in the exchange of GDP with GTP. The Gα subunit releases the bound GDP and binds to GTP, forming a Gα-GTP complex.

3. The Gα-GTP complex dissociates from the Gβγ subunits due to low affinity. The freed complex, then, moves to bind to a specific effector and either activates or inhibits it 2.

4. In this instance, the effector, adenylyl cyclase, is activated by the binding of Gα-GTP complex to convert ATP into cAMP 1. The complex is able to activate or inhibit the effector as long as it remains bound and separated from the Gβγ subunits

1. The ligand binds to the GPCR from the extracellular domain, and induces a conformational change in the receptor on the cytosolic domain 2. The resulting change facilitates the binding of inactive G protein to the receptor.

Figure 2. A diagram showing steps of signal transduction by G protein-coupled receptors.1

Figure 2 (Cont.). A diagram showing steps of signal transduction by G protein-coupled receptors.1

5. The Gα subunit of the complex possesses an endogenous GTPase activity that hydrolyzes GTP into GDP + Pi 2. The hydrolysis typically occurs a few seconds after the complex is bound to the effector. Consequently, GTP (bound to the Gα subunit) is converted to GDP, creating a Gα-GDP complex. 6. The Gα-GDP complex results in a reduction of affinity between the effector and the Gα subunit 2. The complex, thus, dissociates from the effector and rejoins with the Gβγ subunits. Upon the dissociation, the activation or inhibition of the effector ends. 7. Steps 2 to 6 can be repeated since the inactive G protein is reformed and capable of binding to the ligand-bound receptor again. However, the steps

may be halted by a GRK (G proteincoupled receptor kinase) to prevent overstimulation 1. The receptor on the cytosolic domain is phosphorylated by the GRK. 8. The phosphorylated receptor is bound to an arrestin (a molecule that competes for binding with the G protein), which inhibits the GPCR from activating additional G proteins. After a certain period of time, the receptor bound to arrestin will be taken up by endocytosis (a mechanism for the uptake of fluid and solutes into a cell) and replaced with a fresh GPCR to restart the whole process all over again 1.

Summary Signal transduction by G protein-coupled receptors is initiated when the ligand binds to the receptor, altering its conformation and increasing its affinity for the G protein to which it binds. The Gα subunit bound to the GPCR exchanges its GDP with GTP. As a result, the Gα subunit dissociates from the Gβγ subunits and binds to an effector, activating or inhibiting the effector. The activated or inhibited effector produces a respective physiological response. The GTPase activity of the Gα subunit hydrolyzes the bound GTP after a few seconds of binding to the effector, deactivating the Gα subunit. The Gα subunit with GDP reassociates with the Gβγ subunits, ceasing the activity of the effector. The inactive G protein is now reformed and can bind to the receptor again. To avoid overstimulation, however, the receptor is phosphorylated by the GRK and bound by the arrestin, inhibiting additional G proteins from activating. The arrestin-bound receptor is removed after a specific period by endocytosis and replaced with a new GPCR to initiate the whole process all over again. Works Cited 1. Karp G. Cell and Molecular Biology: Concepts and Experiments, 4th ed. Von Hoffmann Press: John Wiley & Sons, Inc; 2005. 2. Compton R. Intracellular Signaling In: BSCI230 Lecture; College Park, MD; 2006 Nov. p. 36-39.