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Application of Site‐Specific Spin Labeling for NMR Detecting Inhibitor‐Induced Conformational Change of HIV‐1 Reverse Transcriptase
Author(s) -
Seetaha Supaporn,
YagiUtsumi Maho,
Yamaguchi Takumi,
Ishii Kentaro,
Hangbua Supa,
Choowongkomon Kiattawee,
Kato Koichi
Publication year - 2016
Publication title -
chemmedchem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.817
H-Index - 100
eISSN - 1860-7187
pISSN - 1860-7179
DOI - 10.1002/cmdc.201500554
Subject(s) - allosteric regulation , chemistry , site directed spin labeling , reverse transcriptase , conformational change , human immunodeficiency virus (hiv) , paramagnetism , chemical shift , drug discovery , mutant , enzyme , nuclear magnetic resonance , biophysics , biochemistry , biology , virology , rna , physics , quantum mechanics , membrane , gene
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.