Collision coupling, crosstalk, and compartmentalization in G-protein coupled receptor systems: can a single model explain disparate results?

The collision coupling model describes interactions between receptors and G-proteins as first requiring the molecules to find each other by diffusion. A variety of experimental data on G-protein activation have been interpreted as suggesting (or not) the compartmentalization of receptors and/or G-proteins in addition to a collision coupling mechanism. In this work, we use a mathematical model of G-protein activation via collision coupling but without compartmentalization to demonstrate that these disparate observations do not imply the existence of such compartments. In experiments with GTP analogs (commonly GTPgammaS), the extent of G-protein activation is predicted to be a function of both receptor number and the rate of GTP analog hydrolysis. The sensitivity of G-protein activation to receptor number is shown to be dependent upon the assay used, with the sensitivity of phosphate production assays (GTPase) >GTPgammaS-binding assays >cAMP inhibition assays. Finally, the amount of competition or crosstalk between receptor species activating the same type of G-proteins is predicted to depend on receptor and G-protein number, but in some (common) experimental regimes this dependence is expected to be minimal. Taken together, these observations suggest that the collision coupling model, without compartments of receptors and/or G-proteins, is sufficient to explain a variety of observations in literature data.

[1]  J. Traynor,et al.  The [35S]GTPgammaS binding assay: approaches and applications in pharmacology. , 2003, Life sciences.

[2]  R. Lefkowitz,et al.  Regulation of G protein-coupled receptor signaling by scaffold proteins. , 2002, Circulation research.

[3]  R. Lefkowitz,et al.  Correlation of beta-adrenergic receptor-stimulated [3H]GDP release and adenylate cyclase activation. Differences between frog and turkey erythrocyte membranes. , 1981, The Journal of biological chemistry.

[4]  R. Pollenz,et al.  Determination of aryl hydrocarbon receptor nuclear translocator protein concentration and subcellular localization in hepatic and nonhepatic cell culture lines: development of quantitative Western blotting protocols for calculation of aryl hydrocarbon receptor and aryl hydrocarbon receptor nuclear t , 1997, Molecular pharmacology.

[5]  J. Linderman,et al.  Threshold and graded response behavior in human neutrophils: effect of varying G-protein or ligand concentrations. , 1998, Biochemistry.

[6]  U. Bhalla,et al.  Emergent properties of networks of biological signaling pathways. , 1999, Science.

[7]  K. Standifer,et al.  Involvement of G Protein-Coupled Receptor Kinase (GRK) 3 and GRK2 in Down-Regulation of the α2B-Adrenoceptor , 2006, Journal of Pharmacology and Experimental Therapeutics.

[8]  J. Traynor,et al.  The [35S]GTPγS binding assay: approaches and applications in pharmacology , 2003 .

[9]  M. Saxton Fluorescence corralation spectroscopy. , 2005, Biophysical journal.

[10]  R. Neubig,et al.  Endogenous RGS Protein Action Modulates μ-Opioid Signaling through Gαo , 2003, The Journal of Biological Chemistry.

[11]  Patricia H Reggio,et al.  Lipids, lipid rafts and caveolae: their importance for GPCR signaling and their centrality to the endocannabinoid system. , 2005, Life sciences.

[12]  R. Neubig,et al.  Compartmentation of receptors and guanine nucleotide-binding proteins in NG108-15 cells: lack of cross-talk in agonist binding among the alpha 2-adrenergic, muscarinic, and opiate receptors. , 1993, Molecular pharmacology.

[13]  R. Neubig,et al.  Endogenous RGS protein action modulates mu-opioid signaling through Galphao. Effects on adenylyl cyclase, extracellular signal-regulated kinases, and intracellular calcium pathways. , 2003, The Journal of biological chemistry.

[14]  J. Traynor,et al.  Endogenous Regulator of G Protein Signaling Proteins Reduce μ-Opioid Receptor Desensitization and Down-Regulation and Adenylyl Cyclase Tolerance in C6 Cells , 2005, Journal of Pharmacology and Experimental Therapeutics.

[15]  A. Yew,et al.  Computational model of the cAMP-mediated sensory response and calcium-dependent adaptation in vertebrate olfactory receptor neurons. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[16]  R. Neubig,et al.  Fluorescence analysis of receptor-G protein interactions in cell membranes. , 2002, Biochemistry.

[17]  J. Traynor,et al.  Relationship between rate and extent of G protein activation: comparison between full and partial opioid agonists. , 2002, The Journal of pharmacology and experimental therapeutics.

[18]  D. Lauffenburger,et al.  Receptors: Models for Binding, Trafficking, and Signaling , 1993 .

[19]  A. Herz,et al.  Opioid receptors are coupled tightly to G proteins but loosely to adenylate cyclase in NG108-15 cell membranes. , 1988, Molecular pharmacology.

[20]  Peter J Woolf,et al.  A Spatial Focusing Model for G Protein Signals , 2003, The Journal of Biological Chemistry.

[21]  D. Lewis,et al.  The CB1 Cannabinoid Receptor Can Sequester G-Proteins, Making Them Unavailable to Couple to Other Receptors , 1999, The Journal of Neuroscience.

[22]  H. Kitano,et al.  A quantitative characterization of the yeast heterotrimeric G protein cycle , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[23]  J. Linderman,et al.  Calculation of diffusion-limited kinetics for the reactions in collision coupling and receptor cross-linking. , 1997, Biophysical journal.

[24]  Terrence P. Kenakin,et al.  A Pharmacologic Analysis of Drug-Receptor Interaction , 1987 .

[25]  E M Ross,et al.  Rapid GTP binding and hydrolysis by G(q) promoted by receptor and GTPase-activating proteins. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[26]  P. Prather,et al.  Chronic exposure to mu-opioid agonists produces constitutive activation of mu-opioid receptors in direct proportion to the efficacy of the agonist used for pretreatment. , 2001, Molecular pharmacology.

[27]  Peter J Woolf,et al.  An algebra of dimerization and its implications for G-protein coupled receptor signaling. , 2004, Journal of theoretical biology.

[28]  Shankar Subramaniam,et al.  Computational modeling reveals how interplay between components of a GTPase-cycle module regulates signal transduction. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[29]  J. Brown,et al.  Differences in muscarinic receptor reserve for inhibition of adenylate cyclase and stimulation of phosphoinositide hydrolysis in chick heart cells. , 1986, Molecular pharmacology.

[30]  A. Levitzki,et al.  Mode of coupling between the beta-adrenergic receptor and adenylate cyclase in turkey erythrocytes. , 1978, Biochemistry.

[31]  M. Millan,et al.  G protein activation by human dopamine D3 receptors in high-expressing Chinese hamster ovary cells: A guanosine-5'-O-(3-[35S]thio)- triphosphate binding and antibody study. , 1999, Molecular pharmacology.

[32]  D. Selley,et al.  Cannabinoid Receptor Agonist Efficacy for Stimulating [35S]GTPγS Binding to Rat Cerebellar Membranes Correlates with Agonist-induced Decreases in GDP Affinity* , 1998, The Journal of Biological Chemistry.

[33]  Akihiro Kusumi,et al.  Detection of non-Brownian diffusion in the cell membrane in single molecule tracking. , 2005, Biophysical journal.

[34]  M. Ui,et al.  [3H]GDP release from rat and hamster adipocyte membranes independently linked to receptors involved in activation or inhibition of adenylate cyclase. Differential susceptibility to two bacterial toxins. , 1984, The Journal of biological chemistry.

[35]  D. Stickle,et al.  Estimation of the kinetic constants for binding of epinephrine to beta-adrenergic receptors of the S49 cell. , 1991, Biochemical pharmacology.

[36]  T. Costa,et al.  Enzymatic degradation of GTP and its "stable" analogues produce apparent isomerization of opioid receptors. , 1989, Journal of receptor research.

[37]  Z. Vogel,et al.  Opioid and Cannabinoid Receptors Share a Common Pool of GTP-Binding Proteins in Cotransfected Cells, But Not in Cells Which Endogenously Coexpress the Receptors , 2000, Cellular and Molecular Neurobiology.

[38]  J. Lameh,et al.  Activity of opioid ligands in cells expressing cloned mu opioid receptors , 2003, BMC pharmacology.

[39]  B. Carter,et al.  Go mediates the coupling of the mu opioid receptor to adenylyl cyclase in cloned neural cells and brain. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[40]  P. Hawkins,et al.  Turkey erythrocyte membranes as a model for regulation of phospholipase C by guanine nucleotides. , 1987, The Journal of biological chemistry.

[41]  J. Linderman,et al.  Compartmentalization of Receptors and Enzymes Affects Activation for a Collision Coupling Mechanism , 1998 .

[42]  J. Woods,et al.  Stimulation of guanosine-5'-o-(3-[35S]thio)triphosphate binding in digitonin-permeabilized C6 rat glioma cells: evidence for an organized association of mu-opioid receptors and G protein. , 2001, The Journal of pharmacology and experimental therapeutics.

[43]  Nicolas Destainville,et al.  Confined diffusion without fences of a g-protein-coupled receptor as revealed by single particle tracking. , 2003, Biophysical journal.

[44]  J. Woods,et al.  Activation of G protein by opioid receptors: role of receptor number and G-protein concentration. , 2000, European journal of pharmacology.

[45]  R. Fantozzi,et al.  Irreversible inactivation of the opiate receptors in the neuroblastoma x glioma hybrid NG108-15 by chlornaltrexamine. , 1981, Molecular Pharmacology.

[46]  J. Woods,et al.  Mu and Delta opioid receptors activate the same G proteins in human neuroblastoma SH‐SY5Y cells , 2002, British journal of pharmacology.

[47]  Rapid kinetics of alpha 2-adrenergic inhibition of adenylate cyclase. Evidence for a distal rate-limiting step. , 1989, Biochemistry.

[48]  Gerda E Breitwieser,et al.  G protein-coupled receptor oligomerization: implications for G protein activation and cell signaling. , 2004, Circulation research.

[49]  N. Gautam,et al.  A Fluorescence Resonance Energy Transfer-based Sensor Indicates that Receptor Access to a G Protein Is Unrestricted in a Living Mammalian Cell*[boxs] , 2004, Journal of Biological Chemistry.

[50]  Y. Sarne,et al.  Divers pathways mediate δ-opioid receptor down regulation within the same cell , 2001 .

[51]  C. Mandyam,et al.  μ-Opioid-Induced Desensitization of Opioid Receptor-Like 1 and μ-Opioid Receptors: Differential Intracellular Signaling Determines Receptor Sensitivity , 2003, Journal of Pharmacology and Experimental Therapeutics.

[52]  R. Rigler,et al.  Fluorescence correlation spectroscopy , 2001 .