Biophysical Studies of G Protein Activation/Deactivation

Summary

Principal Investigator: Richard Neubig
Affiliation: University of Michigan
Country: USA
Abstract: Signal transduction via guanine nucleotide binding proteins (G proteins) is crucial in regulation of cardiovascular, neural, endocrine, and immune cell function as well as in cell growth and differentiation. It has been increasingly recognized that knowledge of the dynamic aspects of signal transduction is important to a full description of signaling mechanisms. This is most evident in the function of the novel family of C protein regulators (RGS proteins). They have been recognized only in the last few years but appear to be responsible for subsecond temporal responses of C protein regulated ion channels and for modulation of slower classical signal transduction pathways. While much in vitro biochemical information is available for RGS proteins, much less is known about their in vivo functions or their potential as targets of drug design. The three aims of this project will: 1) explore mechanisms and specificity of RGS proteins using biochemical and spectroscopic approaches, 2) test via a gene targeting approach the role of RGS proteins in the physiological actions of Gi family G proteins such as regulation of cardiac automaticity and opiate analgesia, tolerance, and dependence, and 3) examine the role and specificity of individual endogenous RGS proteins by use of ribozyme knock-down methods. The studies in this proposal should permit a better understanding of rapid G protein activation and deactivation mechanisms and will explore the potential of RGS proteins as a novel target of drug action.
Funding Period: 1988-12-01 - 2006-02-28
more information: NIH RePORT

Top Publications

  1. ncbi Endogenous RGS proteins and Galpha subtypes differentially control muscarinic and adenosine-mediated chronotropic effects
    Ying Fu
    Department of Pharmacology, University of Michigan, Ann Arbor, MI, USA
    Circ Res 98:659-66. 2006
  2. pmc Isoflurane-induced changes in righting response and breathing are modulated by RGS proteins
    Eduardo E Icaza
    Departments of Anesthesiology, University of Michigan, Ann Arbor, Michigan 48109 5615, USA
    Anesth Analg 109:1500-5. 2009
  3. pmc Design, synthesis and prostate cancer cell-based studies of analogs of the Rho/MKL1 transcriptional pathway inhibitor, CCG-1423
    CHRIS R EVELYN
    Department of Pharmacology, University of Michigan Medical School, University of Michigan, Ann Arbor, MI 48109, USA
    Bioorg Med Chem Lett 20:665-72. 2010
  4. pmc Use of flow cytometric methods to quantify protein-protein interactions
    Levi L Blazer
    Department of Pharmacology, The University of Michigan Medical School, Ann Arbor, Michigan, USA
    Curr Protoc Cytom . 2010
  5. pmc RGS inhibition at G(alpha)i2 selectively potentiates 5-HT1A-mediated antidepressant effects
    Jeffery N Talbot
    Department of Pharmacology, University of Michigan, Ann Arbor, MI 48109, USA
    Proc Natl Acad Sci U S A 107:11086-91. 2010
  6. pmc Gα subunit coordinates with ephrin-B to balance self-renewal and differentiation in neural progenitor cells
    Kiyohito Murai
    Department of Neurosciences, Beckman Research Institute of the City of Hope, Duarte, California 91010, USA
    Stem Cells 28:1581-9. 2010
  7. pmc Thinking outside of the "RGS box": new approaches to therapeutic targeting of regulators of G protein signaling
    Benita Sjögren
    Department of Pharmacology, University of Michigan, 1150 W Medical Center Dr, MSRB III, Ann Arbor, MI 48109, USA
    Mol Pharmacol 78:550-7. 2010
  8. pmc RGS/Gi2alpha interactions modulate platelet accumulation and thrombus formation at sites of vascular injury
    Rachel S Signarvic
    Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA
    Blood 116:6092-100. 2010
  9. pmc Gα(i2)-mediated protection from ischaemic injury is modulated by endogenous RGS proteins in the mouse heart
    Rachael E Waterson
    Department of Pharmaceutical and Biomedical Sciences, Raabe College of Pharmacy, Ohio Northern University, 525 South Main Street, Ada, OH 45810, USA
    Cardiovasc Res 91:45-52. 2011
  10. doi Gαi2 signaling: friend or foe in cardiac injury and heart failure?
    Kuljeet Kaur
    Department of Pharmacology, The University of Michigan Medical School, Ann Arbor, MI 48109 0632, USA
    Naunyn Schmiedebergs Arch Pharmacol 385:443-53. 2012

Detail Information

Publications23

  1. ncbi Endogenous RGS proteins and Galpha subtypes differentially control muscarinic and adenosine-mediated chronotropic effects
    Ying Fu
    Department of Pharmacology, University of Michigan, Ann Arbor, MI, USA
    Circ Res 98:659-66. 2006
    ..Thus, alterations in RGS function may play a role in pathophysiological processes and RGS proteins could represent novel cardiovascular therapeutic targets...
  2. pmc Isoflurane-induced changes in righting response and breathing are modulated by RGS proteins
    Eduardo E Icaza
    Departments of Anesthesiology, University of Michigan, Ann Arbor, Michigan 48109 5615, USA
    Anesth Analg 109:1500-5. 2009
    ....
  3. pmc Design, synthesis and prostate cancer cell-based studies of analogs of the Rho/MKL1 transcriptional pathway inhibitor, CCG-1423
    CHRIS R EVELYN
    Department of Pharmacology, University of Michigan Medical School, University of Michigan, Ann Arbor, MI 48109, USA
    Bioorg Med Chem Lett 20:665-72. 2010
    ..Both compounds were also capable of inhibiting cell invasion with equal efficacy to 1 but with less attendant cytotoxicity...
  4. pmc Use of flow cytometric methods to quantify protein-protein interactions
    Levi L Blazer
    Department of Pharmacology, The University of Michigan Medical School, Ann Arbor, Michigan, USA
    Curr Protoc Cytom . 2010
    ..An adaptation of this method that is compatible for high-throughput screening is also provided...
  5. pmc RGS inhibition at G(alpha)i2 selectively potentiates 5-HT1A-mediated antidepressant effects
    Jeffery N Talbot
    Department of Pharmacology, University of Michigan, Ann Arbor, MI 48109, USA
    Proc Natl Acad Sci U S A 107:11086-91. 2010
    ..By selectively enhancing the beneficial effects of serotonin, inhibition of RGS proteins represents a therapeutic approach for the treatment of mood disorders...
  6. pmc Gα subunit coordinates with ephrin-B to balance self-renewal and differentiation in neural progenitor cells
    Kiyohito Murai
    Department of Neurosciences, Beckman Research Institute of the City of Hope, Duarte, California 91010, USA
    Stem Cells 28:1581-9. 2010
    ....
  7. pmc Thinking outside of the "RGS box": new approaches to therapeutic targeting of regulators of G protein signaling
    Benita Sjögren
    Department of Pharmacology, University of Michigan, 1150 W Medical Center Dr, MSRB III, Ann Arbor, MI 48109, USA
    Mol Pharmacol 78:550-7. 2010
    ..Because several RGS proteins are rapidly degraded by the N-end rule pathway, finding ways to stabilize them may prove to be an effective way to enhance RGS protein function...
  8. pmc RGS/Gi2alpha interactions modulate platelet accumulation and thrombus formation at sites of vascular injury
    Rachel S Signarvic
    Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA
    Blood 116:6092-100. 2010
    ....
  9. pmc Gα(i2)-mediated protection from ischaemic injury is modulated by endogenous RGS proteins in the mouse heart
    Rachael E Waterson
    Department of Pharmaceutical and Biomedical Sciences, Raabe College of Pharmacy, Ohio Northern University, 525 South Main Street, Ada, OH 45810, USA
    Cardiovasc Res 91:45-52. 2011
    ....
  10. doi Gαi2 signaling: friend or foe in cardiac injury and heart failure?
    Kuljeet Kaur
    Department of Pharmacology, The University of Michigan Medical School, Ann Arbor, MI 48109 0632, USA
    Naunyn Schmiedebergs Arch Pharmacol 385:443-53. 2012
    ..Gα(i2) signaling in cardiac myocytes protects against ischemic injury but enhancing Gα(i2) signaling overall may have detrimental effects in heart failure, perhaps through actions on cardiac fibroblasts...
  11. pmc A conserved hydrophobic surface of the LARG pleckstrin homology domain is critical for RhoA activation in cells
    Mohamed Aittaleb
    Life Sciences Institute, University of Michigan, Ann Arbor, MI 48109 2216, USA
    Cell Signal 21:1569-78. 2009
    ..We also show that the hydrophobic patch of the PH domain is likely important for the activity of all Lbc subfamily RhoGEFs...
  12. pmc GNAI2 and regulators of G protein signaling as a potential Noonan syndrome mechanism
    Xinyan Huang
    Department of Pharmacology, University of Michigan, 1301 MSRB III, Ann Arbor, MI 48109, USA
    Med Hypotheses 73:56-9. 2009
    ..This suggests a novel set of candidate genes for NS (GNAI2 and RGS proteins) and if validated could have important implications for therapy as well...
  13. pmc Regulator of G protein signaling protein suppression of Galphao protein-mediated alpha2A adrenergic receptor inhibition of mouse hippocampal CA3 epileptiform activity
    Brianna L Goldenstein
    Department of Pharmacology, Physiology and Therapeutics, School of Medicine and Health Sciences, University of North Dakota, Grand Forks, ND 58202 9037, USA
    Mol Pharmacol 75:1222-30. 2009
    ..This suggests a possible role for RGS inhibitors or selective alpha(2A)AR agonists as a novel antiepileptic drug therapy...
  14. pmc Pleiotropic phenotype of a genomic knock-in of an RGS-insensitive G184S Gnai2 allele
    Xinyan Huang
    Department of Pharmacology, University of Michigan, 1301 MSRB III, Ann Arbor, MI 48109, USA
    Mol Cell Biol 26:6870-9. 2006
    ..Thus, loss of RGS actions at Galpha(i2) produces a dramatic and pleiotropic phenotype which is more evident than the phenotype seen for individual RGS protein knockouts...
  15. ncbi N-terminal residues control proteasomal degradation of RGS2, RGS4, and RGS5 in human embryonic kidney 293 cells
    Johannes Bodenstein
    Department of Pharmacology, 1301 MSRB III Box 0632, 1150 W Medical Center Drive, University of Michigan Medical School, Ann Arbor, MI 48109 0632, USA
    Mol Pharmacol 71:1040-50. 2007
    ..Thus, proteasomal regulation of RGS expression in HEK293 cells strongly controls RGS function and a novel RGS2 mutation with decreased protein expression could be relevant to the pathophysiology of hypertension in humans...
  16. ncbi Endogenous RGS proteins modulate SA and AV nodal functions in isolated heart: implications for sick sinus syndrome and AV block
    Ying Fu
    Department of Pharmacology, University of Michigan, Ann Arbor, MI 48109 0632, USA
    Am J Physiol Heart Circ Physiol 292:H2532-9. 2007
    ..The severe carbachol-induced sinus bradycardia in Galpha(i2)G184S mice suggests a possible role for alterations of Galpha(i2) or RGS proteins in sick sinus syndrome and pathological AV block...
  17. ncbi CCG-1423: a small-molecule inhibitor of RhoA transcriptional signaling
    CHRIS R EVELYN
    Department of Pharmacology, University of Michigan Medical Center, 1301 MSRB III, Room 2220D, 1150 West Medical Center Drive, Ann Arbor, MI 48109 0632, USA
    Mol Cancer Ther 6:2249-60. 2007
    ..Thus, based on its profile, CCG-1423 is a promising lead compound for the development of novel pharmacologic tools to disrupt transcriptional responses of the Rho pathway in cancer...
  18. ncbi Resistance to diet-induced obesity and improved insulin sensitivity in mice with a regulator of G protein signaling-insensitive G184S Gnai2 allele
    Xinyan Huang
    Department of Pharmacology, University of Michigan, 1301 MSRB III, Ann Arbor, MI 48109, USA
    Diabetes 57:77-85. 2008
    ....
  19. pmc Novel peptide ligands of RGS4 from a focused one-bead, one-compound library
    Rebecca A Roof
    Department of Pharmacology, University of Michigan, 1301 MSRB III SPC5632, Ann Arbor, MI 48103, USA
    Chem Biol Drug Des 72:111-9. 2008
    ..Peptide 2 has been modeled to fit in the same binding pocket predicted for YJ34 but in the reverse orientation...
  20. pmc And the winner is ... RGS4!
    Richard R Neubig
    Circ Res 103:444-6. 2008
  21. doi Small molecule protein-protein interaction inhibitors as CNS therapeutic agents: current progress and future hurdles
    Levi L Blazer
    Department of Pharmacology, University of Michigan Medical School, Ann Arbor, MI 48109, USA
    Neuropsychopharmacology 34:126-41. 2009
    ..In particular, we will focus upon recent work towards developing small molecule inhibitors of amyloid-beta and alpha-synuclein aggregation, inhibitors of critical components of G-protein-signaling pathways, and PDZ domain inhibitors...
  22. pmc High-throughput screening for small-molecule inhibitors of LARG-stimulated RhoA nucleotide binding via a novel fluorescence polarization assay
    CHRIS R EVELYN
    Department of Pharmacology, University of Michigan Medical Center, Ann Arbor, Michigan 48109 0632, USA
    J Biomol Screen 14:161-72. 2009
    ..In addition, the fluorescence polarization guanine nucleotide-binding assay described here should serve as a useful approach for both high-throughput screening and general biological applications...
  23. pmc Differential control of opioid antinociception to thermal stimuli in a knock-in mouse expressing regulator of G-protein signaling-insensitive Gαo protein
    Jennifer T Lamberts
    Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA
    J Neurosci 33:4369-77. 2013
    ....