PMID: 7013798Mar 31, 1981Paper

Dihydrofolate reductase hysteresis and its effect of inhibitor binding analyses

Biochemistry
D P Baccanari, S S Joyner

Abstract

Escherichia coli dihydrofolate reductase was shown to follow slow transient kinetics (hysteresis). Nonlinear reaction velocities were detected during the enzyme assay and required 10-15 min to reach a steady-state rate. The degree of hysteresis was influenced by the enzyme concentration and the order of substrate addition. Incubation of the enzyme with NADPH before addition of dihydrofolate resulted in slow initial velocities that increased up to 2-fold during the course of the assay. Increasing the enzyme concentration from 0.2 to 1 nM resulted in diminished hysteresis. NADPH-initiated reactions were linear at all enzyme concentrations tested. Certain drugs had profound effects on hysteresis. Pyrimethamine practically eliminated the hysteresis of dihydrofolate-started reactions, whereas trimethoprime augmented the non-linearities in the sense that hysteresis was detected in both enzyme- and NADPH-started reactions. The shape of these reaction tracings makes trimethoprim is not a slow-binding inhibitor when assayed under conditions that eliminate hysteresis. Contrary to this, sulfamethoxazole did not affect hysteresis or augment inhibition of the enzyme by trimethoprim. Sulfamethoxazole alone (at 6 mM) did not inhibit the hyste...Continue Reading

References

Dec 2, 1975·Biochemistry·D BaccanariJ Burchall
Mar 18, 1977·Proceedings of the Royal Society of London. Series B, Containing Papers of a Biological Character·J FeeneyA Burgen
Nov 1, 1978·Archives of Biochemistry and Biophysics·D P Baccanari
Jun 12, 1979·Biochemistry·J W WilliamsR G Duggleby
Jan 1, 1979·Annual Review of Biochemistry·C Frieden
Feb 14, 1979·Biochemical and Biophysical Research Communications·R E LondonR L Blakley
Jan 1, 1979·Methods in Enzymology·J W Williams, J F Morrison
Aug 1, 1977·Archives of Biochemistry and Biophysics·R C JacksonL I Hart
Dec 1, 1975·Biochemical Pharmacology·S Cha
May 24, 1980·Nucleic Acids Research·D R Smith, J M Calvo
Jan 1, 1980·Advances in Enzyme Regulation·C C FanF M Huennekens

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Citations

Oct 6, 2006·Extremophiles : Life Under Extreme Conditions·Lars RedeckeReinhard Bredehorst
Jul 20, 1994·Biochimica Et Biophysica Acta·S P SassoC M Briand
Jan 13, 2000·European Journal of Biochemistry·V K WalkerR Chaguturu
Aug 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·S L Smith, J J Burchall
Nov 25, 1992·Nucleic Acids Research·E RestoS M Hecht
Aug 5, 2014·Biological & Pharmaceutical Bulletin·Yun-Ju KwonWon-Gon Kim
Jan 17, 2012·Nature Chemical Biology·Mary J CarrollAndrew L Lee
Jan 1, 1986·Scandinavian Journal of Infectious Diseases·L G Burman
Feb 2, 2011·Environmental Microbiology·Gloria Torres-CortésFrancisco Martínez-Abarca
Jul 1, 1984·Biochemical Pharmacology·D P Baccanari, R L Tansik
Feb 8, 1985·The American Journal of Medicine·J D Allan, R C Moellering
Oct 4, 2017·Protein Science : a Publication of the Protein Society·Luis C AcostaAnnelise H Gorensek-Benitez
Apr 7, 2020·Nature Chemistry·Nicole Stéphanie GalenkampGiovanni Maglia
Oct 1, 1996·Antimicrobial Agents and Chemotherapy·T KöhlerJ C Pechere
Nov 18, 1983·Science·J E VillafrancaJ Kraut

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