Infections with the human immunodeficiency virus (HIV-1) and hepatitis C virus (HCV) are among the most significant causes of human morbidity and mortality. Worldwide, there are more than 40 million (HIV-1) and 170 million (HCV) people infected with these viruses. In the United States, 0.6% (HIV-1) and 1.7% (HCV) of the population is infected. More than 25% of those infected with HIV are co-infected with HCV (up to 90% of i.v. drug users). Despite the success of highly active anti-retroviral therapy, there remains a need to develop additional drugs targeted to novel receptors that will ensure different susceptibilities to the development of resistance compared to protease and reverse transcriptase, the principal receptors of current drugs. In contrast, current therapies for HCV are wholly inadequate, and multiple first-generation drugs are needed. Resistance is quickly developed to drugs patterned after those effective against HIV-1. The genomes of both HIV-1 and HCV encode a viroporins, a small membrane protein with ion channel activity involved in the production of new virus particles in infected cells. We propose to take a structural approach to the design of drugs directed against these viroporins. This is technically challenging research because viroporins reside in cell membranes, and one of the most important things we have learned from our studies of membrane proteins is that they are distorted by membrane mimics, such as organic solvents and detergent micelles, and must be studies in their native phospholipid bilayer environment under physiological conditions. As part of this research we have developed a general NMR method for determining the structures of membrane proteins in phospholipid bilayers, and will apply it to these viroporins. Determining the native structures of Vpu (Virus protein "u") from HIV-1 and the p7 protein from HCV are essential in order to understand the molecular mechanism of their biological activities and to accelerate the discovery of drugs that interfere with the activities that contribute to the infectivity of the virses. Because we express these proteins and domains of their cellular partners in bacteria, we have a great deal of flexibility in the design of experiments to characterize their interactions with both drugs and proteins.

Public Health Relevance

Most diseases that afflict humans can be treated or cured with drugs. The majority of therapeutic drugs are chemicals targeted to protein receptors that reside in cell membranes. This research is designed to utilize structural information on the viroporins Vpu from HIV-1 and p7 from HCV to accelerate the discovery of drugs that interfere with their functions.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01GM066978-10
Application #
8711478
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Sakalian, Michael
Project Start
Project End
Budget Start
Budget End
Support Year
10
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of California San Diego
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Das, Bibhuti B; Park, Sang Ho; Opella, Stanley J (2015) Membrane protein structure from rotational diffusion. Biochim Biophys Acta 1848:229-45
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
Lin, Eugene C; Opella, Stanley J (2014) Covariance spectroscopy in high-resolution multi-dimensional solid-state NMR. J Magn Reson 239:57-60
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
Lu, George J; Opella, Stanley J (2014) Mechanism of dilute-spin-exchange in solid-state NMR. J Chem Phys 140:124201
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
Radoicic, Jasmina; Lu, George J; Opella, Stanley J (2014) NMR structures of membrane proteins in phospholipid bilayers. Q Rev Biophys 47:249-83
Das, Bibhuti B; Zhang, Hua; Opella, Stanley J (2014) Dipolar Assisted Assignment Protocol (DAAP) for MAS solid-state NMR of rotationally aligned membrane proteins in phospholipid bilayers. J Magn Reson 242:224-32
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
Das, Bibhuti B; Lin, Eugene C; Opella, Stanley J (2013) Experiments optimized for magic angle spinning and oriented sample solid-state NMR of proteins. J Phys Chem B 117:12422-31

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