G-protein coupled receptors (GPCRs) transduce signals from the outside of cells, by binding to small molecules and their surrogates (drugs), to the inside where they trigger a cascade of events starting with interactions with G-proteins. There are 1000 different GPCRs in human genome, about 450 of which are potential drug targets. More than half of the currently available drugs interact with GPCRs. And since GPCRs are involved in a wide range of biological activities, e.g. blood pressure, pain, cancer growth, etc., the development of new drugs have the potential to cure or ameliorate the symptoms of many diseases. In order to accelerate the rational design of drugs, we propose to develop methods for determining the three dimensional structures of GPCRs. This is a substantial undertaking. On the path towards structure determination we will adapt NMR methods for screening for drug candidates that have been successfully applied to other classes of receptors. This research will be multidisciplinary, involving synthetic organic chemistry, molecular biology, structural biology, NMR spectroscopy, and computer calculations. This research will advance fundamental concepts of protein expression and NMR spectroscopy. It will be highly effective in training scientists that can interact across boundries. It requires the highest levels of technology available; as a result it, involves the University of California, San Diego and two leading biotechnology companies (ProSpect Pharma, Inc., Columbia, MD and m-phasys GmbH, Germany). ProSpect Pharma's key technology is isotopically labeled growth media and their effort is led by Jonathan Miles Brown, Ph.D., COO. M-phasys GmbH has developed methods for expressing GPCRs in bacteria that can utilize ProSpect Pharma's media, and refolding the proteins into their biologically active forms; their effort is led by Hans Kiefer Ph.D., CSO and Stefan Prytulla, Ph.D. GPCRs play a role in a number of human diseases due to their importance in maintaining proper function of living cells. Elucidation of the three-dimensional structures of GPCRs would enable us to look at this family of proteins at the atomic level, aiding in the understanding of how they function as well as how they interact with their respective ligands. This is therefore a very important step in helping to design drugs to target GPCR-related disorders that affect a large number of people including cancer, heart disease and obesity. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
5R01EB005161-02
Application #
7274133
Study Section
Special Emphasis Panel (ZRG1-BCMB-G (90))
Program Officer
Mclaughlin, Alan Charles
Project Start
2006-08-09
Project End
2011-07-31
Budget Start
2007-08-01
Budget End
2008-07-31
Support Year
2
Fiscal Year
2007
Total Cost
$835,151
Indirect Cost
Name
University of California San Diego
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Das, Bibhuti B; Opella, Stanley J (2016) Simultaneous cross polarization to (13)C and (15)N with (1)H detection at 60kHz MAS solid-state NMR. J Magn Reson 262:20-26
Opella, Stanley J (2015) Solid-state NMR and membrane proteins. J Magn Reson 253:129-37
Das, Bibhuti B; Park, Sang Ho; Opella, Stanley J (2015) Membrane protein structure from rotational diffusion. Biochim Biophys Acta 1848:229-45
Opella, Stanley J (2015) Relating structure and function of viral membrane-spanning miniproteins. Curr Opin Virol 12:121-5
Lewinski, Mary K; Jafari, Moein; Zhang, Hua et al. (2015) Membrane Anchoring by a C-terminal Tryptophan Enables HIV-1 Vpu to Displace Bone Marrow Stromal Antigen 2 (BST2) from Sites of Viral Assembly. J Biol Chem 290:10919-33
Park, Sang Ho; Wang, Vivian S; Radoicic, Jasmina et al. (2015) Paramagnetic relaxation enhancement of membrane proteins by incorporation of the metal-chelating unnatural amino acid 2-amino-3-(8-hydroxyquinolin-3-yl)propanoic acid (HQA). J Biomol NMR 61:185-96
Wu, Chin H; De Angelis, Anna A; Opella, Stanley J (2014) Magic angle Lee-Goldburg frequency offset irradiation improves the efficiency and selectivity of SPECIFIC-CP in triple-resonance MAS solid-state NMR. J Magn Reson 246:1-3
Das, Bibhuti B; Opella, Stanley J (2014) Multiple acquisition/multiple observation separated local field/chemical shift correlation solid-state magic angle spinning NMR spectroscopy. J Magn Reson 245:98-104
Tian, Ye; Lu, George J; Marassi, Francesca M et al. (2014) Structure of the membrane protein MerF, a bacterial mercury transporter, improved by the inclusion of chemical shift anisotropy constraints. J Biomol NMR 60:67-71
Tian, Ye; Schwieters, Charles D; Opella, Stanley J et al. (2014) A practical implicit solvent potential for NMR structure calculation. J Magn Reson 243:54-64

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