DOI: 10.1246/bcsj.60.3025. Protein Engineering of Dihydrofolate Reductase. pH Dependency of Phe-31 Mutants

Protein Engineering of Dihydrofolate Reductase. pH Dependency of Phe-31 Mutants

10.1246/bcsj.60.3025

Crossref journal-article

Oxford University Press (OUP)

Bulletin of the Chemical Society of Japan (286)

Abstract

Abstract Site-specific mutations on dihydrofolate reductase from Escherichia coli at the Phe-31 site have generated (Try-31)-DHFR and (Val-31)-DHFR mutant enzymes. The pH dependence of logV and logV⁄KDHF for these enzymes suggests that protonation is important for both the interaction of dihydrofolate and the maximum velocity of the reaction. More importantly, a “hollow” is observed for the Tyr-31 mutant in a logV⁄K–pH profile, necessitating a modification of the wild-type kinetic scheme. The intrinsic pKa of 5.8, obtained based on the modified more general kinetic scheme, for the Tyr-31 mutant agrees well with that obtained from inhibition studies by 2,4-diamino-6,7-dimethylpteridine.

Bibliography

Taira, K., Chen, J.-T., Fierke, C. A., & Benkovic, S. J. (1987). Protein Engineering of Dihydrofolate Reductase. pH Dependency of Phe-31 Mutants. Bulletin of the Chemical Society of Japan, 60(8), 3025â3030.

Authors 4

Kazunari Taira (first)
Jin-Tann Chen (additional)
Carol A Fierke (additional)
Stephen J Benkovic (additional)

References 17 Referenced 5

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10.1002/jcb.240290203
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10.1246/bcsj.60.3017
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10.1021/bi00307a034
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10.1016/0167-4838(83)90055-9
{'key': '2024012102070970800_r16'}
10.1126/science.3511529

Dates

Type	When
Created	19 years ago (July 25, 2006, 9:45 p.m.)
Deposited	1 year, 7 months ago (Jan. 20, 2024, 9:07 p.m.)
Indexed	1 year, 7 months ago (Jan. 21, 2024, 12:01 a.m.)
Issued	38 years ago (Aug. 1, 1987)
Published	38 years ago (Aug. 1, 1987)
Published Online	19 years, 1 month ago (June 27, 2006)
Published Print	38 years ago (Aug. 1, 1987)

Funders 0

None

BibTeX

@article{Taira_1987, title={Protein Engineering of Dihydrofolate Reductase. pH Dependency of Phe-31 Mutants}, volume={60}, ISSN={1348-0634}, url={http://dx.doi.org/10.1246/bcsj.60.3025}, DOI={10.1246/bcsj.60.3025}, number={8}, journal={Bulletin of the Chemical Society of Japan}, publisher={Oxford University Press (OUP)}, author={Taira, Kazunari and Chen, Jin-Tann and Fierke, Carol A and Benkovic, Stephen J}, year={1987}, month=aug, pages={3025–3030} }

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