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  Kinetics of interaction of HIV reverse transcriptase with primer/template

Divita, G., Mueller, B., Immendörfer, U., Gautel, M., Rittinger, K., Restle, T., et al. (1993). Kinetics of interaction of HIV reverse transcriptase with primer/template. Biochemistry, 32(31), 7966-7971. doi:10.1021/bi00082a018.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0019-AA1C-2 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002D-2622-B
Genre: Journal Article

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 Creators:
Divita, Gilles, Author
Mueller, B., Author
Immendörfer, Ulrike1, Author              
Gautel, Mathias, Author
Rittinger, Katrin1, Author              
Restle, Tobias1, 2, 3, Author              
Goody, Roger S.1, Author              
Affiliations:
1Emeritus Group Biophysics, Max Planck Institute for Medical Research, Max Planck Society, ou_1497712              
2Molecular chaperones, Max Planck Institute for Medical Research, Max Planck Society, ou_1497728              
3Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Max Planck Society, ou_1497700              

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 Abstract: Intrinsic protein fluorescence of reverse transcriptases from HIV-1 and HIV-2 provides a sensitive signal for monitoring the interaction of the enzymes with primer/template duplex molecules. Kd values for 18/36-mer DNA/DNA duplexes were found to be in the range of a few nanomolar (about 3 times higher for the enzyme from HIV-2 than for that from HIV-1). The quenching of protein fluorescence induced on binding primer/template, together with an increase in extrinsic fluorescence on interaction with primer/template containing a fluorescent nucleotide at the 3'-end of the primer, was used to investigate the kinetics of interaction with reverse transcriptase from HIV-1. The results can be explained in terms of a two-step binding model, with a rapid diffusion-limited initial association (k(ass) = ca. 5 x 10(8) M-1 s-1) followed by a slow isomerization step (k = ca. 0.5 s-1). These (forward) rate constants are increased in the presence of a non-nucleoside inhibitor (S-TIBO) of HIV-1 reverse transcriptase, while the reverse rate constant for the second step is decreased, leading to an increase in affinity between the enzyme and primer/template by a factor of at least 10 when S-TIBO is bound. The results are discussed in terms of present knowledge of the structure of reverse transcriptase.

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Language(s): eng - English
 Dates: 1993-03-041993-04-211993-08-10
 Publication Status: Published in print
 Pages: 6
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 Rev. Method: Peer
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Title: Biochemistry
Source Genre: Journal
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Publ. Info: Columbus, Ohio : American Chemical Society
Pages: - Volume / Issue: 32 (31) Sequence Number: - Start / End Page: 7966 - 7971 Identifier: ISSN: 0006-2960
CoNE: https://pure.mpg.de/cone/journals/resource/954925384103