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Temperature-dependent equilibrium between the open and closed conformation of the p66 subunit of the HIV-1 reverse transcriptase revealed by site-directed spin labelling

Title data

Kensch, Oliver ; Restle, Tobias ; Wöhrl, Birgitta ; Goody, Roger S. ; Steinhoff, Heinz-Jürgen:
Temperature-dependent equilibrium between the open and closed conformation of the p66 subunit of the HIV-1 reverse transcriptase revealed by site-directed spin labelling.
In: Journal of Molecular Biology. Vol. 301 (2000) Issue 4 . - pp. 1029-1039.
ISSN 0022-2836
DOI: https://doi.org/10.1006/jmbi.2000.3998

Abstract in another language

X-ray crystallographic studies of human immunodeficiency virus type 1 reverse transcriptase complexed with or without substrates or inhibitors show that the heterodimeric enzyme adopts distinct conformations that differ in the orientation of the so-called thumb subdomain in the large subunit. Site-directed spin labelling of mutated residue positions W24C and K287C is applied here to determine the distances between the fingers and thumb subdomains of liganded and unliganded RT in solution. The inter-spin distances of a DNA/DNA and a pseudoknot RNA complexed reverse transcriptase in solution was found to agree with the respective crystal data of the open and closed conformations. For the unliganded reverse transcriptase a temperature-dependent equilibrium between these two states was observed. The fraction of the closed conformation decreased from 95 % at 313 K to 65 % at 273 K. The spectral separation between the two structures was facilitated by the use of a perdeuterated [¹⁵N]nitroxide methane-thiosulfonate spin label.

Further data

Item Type: Article in a journal
Refereed: Yes
Institutions of the University: Faculties > Faculty of Biology, Chemistry and Earth Sciences
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biopolymers
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biopolymers > Lehrstuhl Biopolymere - Apl. Prof. Dr. Birgitta Wöhrl
Result of work at the UBT: No
DDC Subjects: 500 Science > 540 Chemistry
500 Science > 570 Life sciences, biology
Date Deposited: 17 May 2019 07:33
Last Modified: 17 May 2019 07:33
URI: https://eref.uni-bayreuth.de/id/eprint/48976