DOI: 10.1098/rstb.2006.1871. The role of enzyme dynamics and tunnelling in catalysing hydride transfer: studies of distal mutants of dihydrofolate reductase

The role of enzyme dynamics and tunnelling in catalysing hydride transfer: studies of distal mutants of dihydrofolate reductase

10.1098/rstb.2006.1871

Crossref journal-article

The Royal Society

Philosophical Transactions of the Royal Society B: Biological Sciences (175)

Abstract

Residues M42 and G121 of Escherichia coli dihydrofolate reductase ( ec DHFR) are on opposite sides of the catalytic centre (15 and 19 Å away from it, respectively). Theoretical studies have suggested that these distal residues might be part of a dynamics network coupled to the reaction catalysed at the active site. The ec DHFR mutant G121V has been extensively studied and appeared to have a significant effect on rate, but only a mild effect on the nature of H-transfer. The present work examines the effect of M42W on the physical nature of the catalysed hydride transfer step. Intrinsic kinetic isotope effects (KIEs), their temperature dependence and activation parameters were studied. The findings presented here are in accordance with the environmentally coupled hydrogen tunnelling. In contrast to the wild-type (WT), fluctuations of the donor–acceptor distance were required, leading to a significant temperature dependence of KIEs and deflated intercepts. A comparison of M42W and G121V to the WT enzyme revealed that the reduced rates, the inflated primary KIEs and their temperature dependences resulted from an imperfect potential surface pre-arrangement relative to the WT enzyme. Apparently, the coupling of the enzyme's dynamics to the reaction coordinate was altered by the mutation, supporting the models in which dynamics of the whole protein is coupled to its catalysed chemistry.

Bibliography

Wang, L., Goodey, N. M., Benkovic, S. J., & Kohen, A. (2006). The role of enzyme dynamics and tunnelling in catalysing hydride transfer: studies of distal mutants of dihydrofolate reductase. Philosophical Transactions of the Royal Society B: Biological Sciences, 361(1472), 1307â1315.

Authors 4

Lin Wang (first)
Nina M Goodey (additional)
Stephen J Benkovic (additional)
Amnon Kohen (additional)

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Dates

Type	When
Created	19 years, 1 month ago (July 13, 2006, 4:06 p.m.)
Deposited	4 years, 6 months ago (Feb. 20, 2021, 9:40 p.m.)
Indexed	4 months, 1 week ago (April 29, 2025, 10:05 a.m.)
Issued	19 years, 1 month ago (July 12, 2006)
Published	19 years, 1 month ago (July 12, 2006)
Published Online	19 years, 1 month ago (July 12, 2006)
Published Print	19 years ago (Aug. 29, 2006)

Funders 0

None

BibTeX

@article{Wang_2006, title={The role of enzyme dynamics and tunnelling in catalysing hydride transfer: studies of distal mutants of dihydrofolate reductase}, volume={361}, ISSN={1471-2970}, url={http://dx.doi.org/10.1098/rstb.2006.1871}, DOI={10.1098/rstb.2006.1871}, number={1472}, journal={Philosophical Transactions of the Royal Society B: Biological Sciences}, publisher={The Royal Society}, author={Wang, Lin and Goodey, Nina M and Benkovic, Stephen J and Kohen, Amnon}, year={2006}, month=jul, pages={1307–1315} }

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