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Characterization and inhibition of dihydrofolate synthetase from Neisseria gonorrhoeae

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Summary

Dihydrofolate synthetase (EC 6.3.2.12) from N. gonorrhoeae was isolated and enzyme characteristics were determined. The purified enzyme was found quite stable when stored at −60 °C. About 50% of the enzyme activity wag destroyed within 6 weeks when kept at 4 °C. Maximum velocity was observed at pH 9.3. The enzyme required a monovalent cation, K+ or NH4 + , and divalent cation, Mg2+ or Mn2+ for its function. ATP at 5 mM concentration gave maximum activity. Km values for dihydropteroate and L-glutamate at pH 9.3 were 3.5 × 10−5 M and 6.5 × 10−4 M, respectively. Patterns of product inhibition by dihydrofolate were found to be non-competitive with respect to dihydropteroate, having a Ki value of 5.1 ± 0.8 × 10−4 M, and competitive with respect to L-glutamate, having a Ki value of 6.2 × 10−4 M.

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Authors and Affiliations

  1. Samuel M. Best Research Laboratory, Massachusetts College of Pharmacy and Allied Health Sciences, 02115, Boston, MA, USA

    Sunanta Pongsamart, Richard I. Ho, Leonard Corman & William O. Foye

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  1. Sunanta Pongsamart
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  2. Richard I. Ho
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  3. Leonard Corman
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  4. William O. Foye
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Pongsamart, S., Ho, R.I., Corman, L. et al. Characterization and inhibition of dihydrofolate synthetase from Neisseria gonorrhoeae . Mol Cell Biochem 59, 165–171 (1984). https://doi.org/10.1007/BF00231312

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  • DOI: https://doi.org/10.1007/BF00231312

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