1. Application of Site-Specific Spin Labeling for NMR Detecting Inhibitor-Induced Conformational Change of HIV-1 Reverse Transcriptase
- Author
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Takumi Yamaguchi, Supaporn Seetaha, Koichi Kato, Kentaro Ishii, Maho Yagi-Utsumi, Supa Hannongbua, and Kiattawee Choowongkomon
- Subjects
0301 basic medicine ,Models, Molecular ,Conformational change ,Protein Conformation ,Allosteric regulation ,HIV Infections ,Nuclear Overhauser effect ,010402 general chemistry ,01 natural sciences ,Biochemistry ,03 medical and health sciences ,Drug Discovery ,Humans ,Carbon Radioisotopes ,General Pharmacology, Toxicology and Pharmaceutics ,Nuclear Magnetic Resonance, Biomolecular ,Pharmacology ,Drug discovery ,Chemistry ,Organic Chemistry ,Nuclear magnetic resonance spectroscopy ,Site-directed spin labeling ,Molecular biology ,Reverse transcriptase ,HIV Reverse Transcriptase ,0104 chemical sciences ,030104 developmental biology ,Biophysics ,HIV-1 ,Molecular Medicine ,Reverse Transcriptase Inhibitors ,Target protein - Abstract
Paramagnetism-assisted nuclear magnetic resonance (NMR) techniques can provide long-range structural information complemented with local information derived from chemical-shift perturbation and nuclear Overhauser effect data. Here, we address the application of paramagnetic relaxation enhancement (PRE) to detect inhibitor-induced conformational change of a drug target protein using human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) as a model protein. Using a site-specific spin-labeled HIV-1 RT mutant with selective (13) C labeling, conformation-dependent PREs were successfully observed reflecting the stabilization of an open conformation of this enzyme caused by inhibitor binding. This study demonstrates that the paramagnetism-assisted NMR approach offers an alternative strategy in protein-based drug screening to identify allosteric inhibitors of a target protein.
- Published
- 2015