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G-Protein-Coupled Receptor Heteromer Dynamics

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G-Protein-Coupled Receptor Heteromer Dynamics Commentary 4215 G-protein-coupled receptor heteromer dynamics Jean-Pierre Vilardaga1,2,*, Luigi F. Agnati3, Kjell Fuxe4 and Francisco Ciruela5 1Laboratory for GPCR Biology, Department of Pharmacology and Chemical Biology, University of Pittsburgh, School of Medicine, Pittsburgh, PA 15261, USA 2Endocrine Unit, Department of Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA 3IRCCS, San Camillo, Lido Venezia, Italy 4Department of Neuroscience, Karolinska Institute, Stockholm SE-17177, Sweden 5Pharmacology Unit, Department of Pathology and Experimental Therapy, Faculty of Medicine, University of Barcelona, 08907 Barcelona, Spain *Author for correspondence ([email protected]) Journal of Cell Science 123, 000-000 © 2010. Published by The Company of Biologists Ltd doi:10.1242/jcs.063354 Summary G-protein-coupled receptors (GPCRs) represent the largest family of cell surface receptors, and have evolved to detect and transmit a large palette of extracellular chemical and sensory signals into cells. Activated receptors catalyze the activation of heterotrimeric G proteins, which modulate the propagation of second messenger molecules and the activity of ion channels. Classically thought to signal as monomers, different GPCRs often pair up with each other as homo- and heterodimers, which have been shown to modulate signaling to G proteins. Here, we discuss recent advances in GPCR heteromer systems involving the kinetics of the early steps in GPCR signal transduction, the dynamic property of receptor–receptor interactions, and how the formation of receptor heteromers modulate the kinetics of G-protein signaling. Key words: G-protein-coupled receptors, Heterodimers, Signaling Introduction the signaling and trafficking mechanisms of GPCRs is thus central G-protein-coupled receptors (GPCRs) constitute the main family for the development of new and safer therapies for many of cell surface receptors for a large variety of chemical stimuli physiological and psychological disorders. (hormones, neurotransmitters, chemoattractants, calcium ions and GPCRs have traditionally been considered to exist and function pain killers, among others molecules) and sensory stimuli (light, as cell-surface-receptor monomers (Chabre and Le Maire, 2005; odorants and taste molecules). GPCRs are expressed in virtually Chabre et al., 2009). Consistent with this view, recent studies every cell and consist of seven membrane-spanning a-helical characterized in reconstituted phospholipid bilayer systems structures with the N- and C-termini exposed to the extracellular demonstrated that purified receptors such as rhodopsin, b2-adrenergic Journal of Cell Science and intracellular environment, respectively (Hanson and Stevens, and -opioid receptors can activate heterotrimeric G proteins as 2009). Signal transduction begins when an extracellular agonist single GPCR monomers (Whorton et al., 2007; Whorton et al., ‘ligand’ binds and switches the receptor from an inactive state to 2008; Kuszak, 2008). These studies indicate that a single GPCR an active state conformation. Activated receptors then catalyze the protomer is capable of activating G proteins, and that receptor exchange of GDP for GTP on the a-subunit of heterotrimeric G oligomerization is not necessary to mediate G-protein activation. proteins (Gabg), which in turn engages conformational and/or Several GPCRs can, however, assemble and signal in both homo- dissociation events between the Ga and dimeric Gbg subunits that oligomer and hetero-oligomer complexes at the surface membrane are associated with G-protein activation (Hofmann et al., 2009). of native and cultured cells (Prinster et al., 2005; Guo et al., 2008; Both the GTP-bound Ga subunit and the Gbg dimer can then Milligan, 2009) (Box 1). Two recent studies provided strong initiate or suppress the activity of effector enzymes (e.g. adenylyl experimental evidence of the in vivo homodimerization of GPCRs. cyclases, phosphodiesterases, phospholipases) and ion channels. The first showed dimerization of the mouse luteinizing hormone These ion channels in turn modulate the flow of secondary receptor (LHR) by using a transgenetic approach to express binding messengers such as cAMP, cGMP, diacylglycerol or inositol and signaling variants of LHR together (Rivero-Muller et al., 2010), trisphosphate, which are involved in the regulation of multiple whereas the second study demonstrated the existence of native intracellular signaling pathways, and which modulate cell functions oxytocin receptor dimers by using a fluorescence resonance energy in body systems as diverse as the skeletal, endocrine, cardiovascular transfer (FRET) approach (Albizu et al., 2010). In this Commentary, and nervous systems, among others (Fig. 1). Activated GPCRs can we discuss the kinetics of the early steps involved in GPCR signaling also interact with cytosolic arrestins (b-arrestin-1 and b-arrestin- (receptor activation, the receptor–G-protein interaction, G-protein 2), which coordinate receptor–G-protein uncoupling in space and activation), the stability of the receptor–receptor interaction, and time, promote receptor internalization and modulate various distal how this interaction modulates the kinetics of G-protein signaling. signaling responses such the MAP kinase cascades (Pierce et al., 2002). Malfunction of GPCRs are involved in many human Initial steps in the GPCR signaling system diseases (Schöneberg et al., 2004), for instance, diabetes insipidus Biophysical and biochemical approaches have revealed that GPCR and mellitus, hypercalcemia, obesity, hypertension, cancer, signaling systems involve a succession of events that initially take hypothyroidism, retinitis pigmentosa and psychotic disorders, and place at the cell membrane and modulate the production and are targets of most of the available clinical drugs used in humans, propagation of second-messenger molecules inside the cell (Fig. 2) such as b-blockers, antipsychotics and analgesics. Understanding (Lamb, 1996; Ferrandon et al., 2009). These events are initiated by 4216 Journal of Cell Science 123 (24) particularly helices 3 and 6 (Bourne, 1997). Activation of the b2- Box 1. Receptor heterodimer and receptor mosaic adrenergic receptor (b2-AR), for example, proceeds through the concepts release of an ‘ionic lock’ that holds together the cytoplasmic sides In the 1980s, Agnati and Fuxe introduced the concept of of helix 3 and helix 6 in the inactive conformation of the receptor, intermembrane receptor–receptor interactions between different and a rotamer toggle switch mechanism that modulates the types of GPCRs, based on the effects of neuropeptides on the binding characteristics of monoamine receptors in plasma conformation of helix 6 (Yao et al., 2006). These conformational membrane preparations from discrete regions of the brain (Agnati changes are considered to expose the intracellular receptor domains et al., 1980; Fuxe et al., 1981; Fuxe et al., 1983; Fuxe and Agnati, at the cytosolic side, which interact and activate G proteins. Our 1985). These results were in line with earlier findings (Limbird et FRET approach (for a review, see Vilardaga et al., 2009; Lohse al., 1975) showing negative cooperativity in b-ARs, which could et al., 2008) revealed that intramolecular rearrangements associated be explained by the existence of receptor homodimers leading to with receptor activation in live cells proceed with fast kinetics site–site interactions. Additional in vivo studies, which examined (Vilardaga et al., 2003). The activation of a receptor by a small the interactions of clonidine (a partial agonist for a -ARs) and 2A neurotransmitter such as the adenosine A2A receptor, a2A- and neuropeptide Y (NPY; a 36 amino acid neurotransmitter) on b1-adrenergic receptors, and muscarinic receptors takes place with sleep–wakefulness cycles and arterial blood pressure control in a time constant tϷ40 mseconds, whereas activation of the receptor both normal and spontaneously hypertensive rats (Fuxe et al., Ϸ 1989; Fuxe, 1990; Yang et al., 1994), further supported the for the larger parathyroid hormone proceeds within t 1 second existence of receptor–receptor interactions and their potential (Vilardaga et al., 2003; Vilardaga, 2010; Maier-Peuschel et al., functional relevance. As a logical consequence for the indications 2010). One of the mechanistic reasons for this 25-fold difference of direct physical interactions between neuropeptide and in receptor activation kinetics is thought to depend on distinct monoamine receptors, the term heteromerization was introduced modes of binding between peptide hormones and smaller molecules to describe a specific interaction between different types of (Castro et al., 2005; Vilardaga, 2010). GPCRs (Zoli et al., 1993). The concept of the GPCR heterodimer Activated GPCRs then interact with heterotrimeric G proteins was later confirmed by studies reporting that two non-functional with kinetics that, in the context of the low level of G proteins, are GPCR monomers, GABAB1 and GABAB2, can assemble in not determined by the time course of receptor activation, but rather signaling heterodimers at the cell surface to transmit the by a diffusion-limited collision process. At a high level of G- inhibitory effect of the g-aminobutyric acid (GABA) neurotransmitter (Marshall et al., 1999): when expressed alone, protein expression, however, a receptor–G-protein interaction can be as fast as receptor activation itself, with a time constant ranging the GABAB1 receptor is retained in
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