title: Mass Spectrometric Characterization of HIV-1 Reverse Transcriptase Interactions with Non-nucleoside Reverse Transcriptase Inhibitors.
abstract: Non-nucleoside reverse transcriptase inhibitors (NNRTIs) of human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) have been developed for the treatment of acquired immunodeficiency syndrome. HIV-1 RT binding to NNRTIs has been characterized by various biophysical techniques. However, these techniques are often hampered by the low water solubility of the inhibitors, such as the current promising diarylpyrimidine-based inhibitors rilpivirine and etravirine. Hence, a conventional and rapid method that requires small sample amounts is desirable for studying NNRTIs with low water solubility. Here we successfully applied a recently developed mass spectrometric technique under non-denaturing conditions to characterize the interactions between the heterodimeric HIV-1 RT enzyme and NNRTIs with different inhibitory activities. Our data demonstrate that mass spectrometry serves as a semi-quantitative indicator of NNRTI binding affinity for HIV-1 RT using low and small amounts of samples, offering a new high-throughput screening tool for identifying novel RT inhibitors as anti-HIV drugs.
authors.affiliation: Department of Chemistry, Faculty of Science, Kasetsart University.
authors.forename: Ratsupa
authors.initials: R
authors.lastname: Thammaporn
authors.forename: Kentaro
authors.initials: K
authors.lastname: Ishii
authors.forename: Maho
authors.initials: M
authors.lastname: Yagi-Utsumi
authors.forename: Susumu
authors.initials: S
authors.lastname: Uchiyama
authors.forename: Supa
authors.initials: S
authors.lastname: Hannongbua
authors.forename: Koichi
authors.initials: K
authors.lastname: Kato
mesh_terms: D000480:Alkynes; D019380:Anti-HIV Agents / Q000378:metabolism*; D048588:Benzoxazines / Q000378:metabolism; D003521:Cyclopropanes; D054303:HIV Reverse Transcriptase / Q000378:metabolism*; D013058:Mass Spectrometry; D019829:Nevirapine / Q000378:metabolism; D009570:Nitriles; D011724:Pyridazines / Q000378:metabolism; D011743:Pyrimidines; D018894:Reverse Transcriptase Inhibitors / Q000378:metabolism*; D000068696:Rilpivirine / Q000378:metabolism
chemical_list: D000480:Alkynes; D019380:Anti-HIV Agents; D048588:Benzoxazines; D003521:Cyclopropanes; D009570:Nitriles; D011724:Pyridazines; D011743:Pyrimidines; D018894:Reverse Transcriptase Inhibitors; C451734:etravirine; D019829:Nevirapine; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase; D000068696:Rilpivirine; C098320:efavirenz