NMR characterization of HIV-1 reverse transcriptase binding to various non-nucleoside reverse transcriptase inhibitors with different activities.
Human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) is an important target for antiviral therapy against acquired immunodeficiency syndrome. However, the efficiency of available drugs is impaired most typically by drug-resistance mutations in this enzyme. In this study, we applied a nuclear magnetic resonance (NMR) spectroscopic technique to the characterization of the binding of HIV-1 RT to various non-nucleoside reverse transcriptase inhibitors (NNRTIs) with different activities, i.e., nevirapine, delavirdine, efavirenz, dapivirine, etravirine, and rilpivirine. (1)H-(13)C heteronuclear single-quantum coherence (HSQC) spectral data of HIV-1 RT, in which the methionine methyl groups of the p66 subunit were selectively labeled with (13)C, were collected in the presence and absence of these NNRTIs. We found that the methyl (13)C chemical shifts of the M230 resonance of HIV-1 RT bound to these drugs exhibited a high correlation with their anti-HIV-1 RT activities. This methionine residue is located in proximity to the NNRTI-binding pocket but not directly involved in drug interactions and serves as a conformational probe, indicating that the open conformation of HIV-1 RT was more populated with NNRTIs with higher inhibitory activities. Thus, the NMR approach offers a useful tool to screen for novel NNRTIs in developing anti-HIV drugs.
Department of Chemistry, Faculty of Science, Kasetsart University, Chatuchak, Bangkok, Thailand.|Institute for Molecular Science and Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences, Japan.
Ratsupa
R
Thammaporn
Institute for Molecular Science and Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences, Japan.|Graduate School of Pharmaceutical Sciences, Nagoya City University, Japan.
Maho
M
Yagi-Utsumi
Institute for Molecular Science and Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences, Japan.|Graduate School of Pharmaceutical Sciences, Nagoya City University, Japan.
Takumi
T
Yamaguchi
Department of Chemistry, Faculty of Science, Srinakharinwirot University, Bangkok, Thailand.
Pornthip
P
Boonsri
Department of Chemistry, Faculty of Science, Kasetsart University, Chatuchak, Bangkok, Thailand.|Center of Nanotechnology, Kasetsart University, Chatuchak, Bangkok, Thailand.
Patchreenart
P
Saparpakorn
Department of Biochemistry, Faculty of Science, Kasetsart University, Chatuchak, Bangkok, Thailand.
Kiattawee
K
Choowongkomon
Department of Chemistry, Faculty of Science, Kasetsart University, Chatuchak, Bangkok, Thailand.
Supanna
S
Techasakul
Institute for Molecular Science and Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences, Japan.|Graduate School of Pharmaceutical Sciences, Nagoya City University, Japan.
Koichi
K
Kato
Department of Chemistry, Faculty of Science, Kasetsart University, Chatuchak, Bangkok, Thailand.|Center of Nanotechnology, Kasetsart University, Chatuchak, Bangkok, Thailand.
Supa
S
Hannongbua
D001665:Binding Sites; D054303:HIV Reverse Transcriptase / Q000037:antagonists & inhibitors / Q000737:chemistry*; D015497:HIV-1 / Q000201:enzymology*; D006801:Humans; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D018894:Reverse Transcriptase Inhibitors / Q000737:chemistry*
D018894:Reverse Transcriptase Inhibitors; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase