Biosensor-based kinetic characterization of the interaction between HIV-1 reverse transcriptase and non-nucleoside inhibitors

• Details of the interaction between HIV-1 reverse transcriptase and non-nucleoside inhibitors (NNRTIs) have been elucidated using a biosensor-based approach. This initial study was performed with HIV-1 reverse transcriptase mutant K103N, the phenethylthioazolylthiourea compound (PETT) MIV-150, and the three NNRTIs licensed for clinical use: nevirapine, delavirdine, and efavirenz. Mathematical evaluation of the experimental data with several interaction models revealed that the four inhibitors interacted with HIV-1 RT with varying degrees of complexity. The simplest adequate model accounted for two different conformations of the free enzyme, of which only one can bind the inhibitor, consistent with a previously hypothesized population-shift model including a preformation of the NNRTI binding site. In addition, a heterogeneous binding was observed for delavirdine, efavirenz, and MIV-150, indicating that two noncompetitive and kinetically distinct enzyme-inhibitor complexes could be formed. Furthermore, for these compounds, there were indications for ligand-induced conformational changes.

Ämnesord

NATURVETENSKAP  -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)

Nyckelord

Binding Sites

Biosensing Techniques/*methods

Delavirdine/chemistry/pharmacology

Enzyme Activation/drug effects

HIV-1 Reverse Transcriptase/*antagonists & inhibitors/chemistry

Kinetics

Molecular Structure

Nevirapine/chemistry/pharmacology

Oxazines/chemistry/pharmacology

Protein Conformation

Pyridines/chemistry/pharmacology

Reverse Transcriptase Inhibitors/chemistry/*pharmacology

Structure-Activity Relationship

Time Factors

Urea/analogs & derivatives/chemistry/pharmacology

Cell and molecular biology

Cell- och molekylärbiologi

Publikations- och innehållstyp

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