Research article summary (published 27 Sep 2009):

Kinetics of association and dissociation of HIV-1 reverse transcriptase subunits.

Full Abstract

The biologically active form of HIV-1 reverse transcriptase (RT) is the p66/p51 heterodimer. The process of maturation of the heterodimer from precursor proteins is poorly understood. Previous studies indicated that association of p66 and p51 is very slow. Three techniques, a pre-steady-state activity assay, intrinsic tryptophan fluorescence, and a FRET assay, were used to monitor the dimerization kinetics of RT. Kinetic experiments were conducted with purified p66 and p51 proteins in aqueous buffer. All three techniques gave essentially the same results. The dissociation kinetics of p66/p51 were first-order with rate constants (k(diss)) of approximately 4 x 10(-6) s(-1) (t(1/2) = 48 h). The association kinetics of p66 and p51 were concentration-dependent with second-order rate constants (k(ass)) of approximately 1.7 M(-1) s(-1) for the simple bimolecular association reaction. The implications of slow dimerization of p66/p51 for the maturation process are discussed. A reaction-controlled model invoking conformational selection is proposed to explain the slow protein-protein association kinetics.

Author information

Author/s: Venezia, Carl F (CF); Meany, Brendan J (BJ); Braz, Valerie A (VA); Barkley, Mary D (MD);

Affiliation: Department of Chemistry, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, Ohio 44106, USA.

Grants: GM071267 (Agency:NIGMS NIH HHS) ; HL07653 (Agency:NHLBI NIH HHS)

Journal and publication information

Publication Type: Journal Article; Research Support, N.I.H., Extramural

Journal: Biochemistry (Biochemistry), published in United States. (Language: eng)

Reference: 2009-Sep; vol 48 (issue 38) : pp 9084-93

Dates: Created 2009/09/22; Completed 2009/10/09; Revised 2009/11/02;

PMID: 19715314, status: MEDLINE (last retrieval date: 11/3/2009, IMS Date: )

Sourced from the National Library of Medicine. Abstract text and other information may be subject to copyright.

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MeSH headings (categories)

This article was linked to the MESH Headings shown below.

Amino Acid Substitution Base Sequence DNA Primers - genetics Dimerization Fluorescence Resonance Energy Transfer HIV Reverse Transcriptase - chemistry; genetics; metabolism HIV-1 - enzymology; genetics Kinetics

Models, Molecular Mutagenesis, Site-Directed Protein Structure, Quaternary Protein Subunits Recombinant Proteins - chemistry; genetics; metabolism Spectrometry, Fluorescence Thermodynamics Tryptophan - chemistry

Associated Chemicals: DNA Primers (0) ; Protein Subunits (0) ; Recombinant Proteins (0) ; Tryptophan (73-22-3) ; reverse transcriptase, Human immunodeficiency virus 1 (EC 2.7.7.-) ; HIV Reverse Transcriptase (EC 2.7.7.49)

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