Formyl phosphate: A proposed intermediate in the reaction catalyzed by Escherichia coli PurT GAR transformylase

Ariane E. Marolewski, Karen M. Mattia, Mark S. Warren, Stephen J. Benkovic

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

The Escherichia coli purT encoded glycinamide ribonucleotide transformylase (GAR transformylase) serves as an alternate enzyme in the production of formyl GAR for use in de novo purine biosynthesis. This enzyme differs from the previously known purN encoded enzyme in size, sequence, and substrates; ATP and formate are required as opposed to formyl tetrahydrofolate. Kinetic studies of the wild-type PurT enzyme described here demonstrate that formyl phosphate behaves as a chemically and kinetically competent intermediate. The requirement for ATP and GAR in these reactions is consistent with previous steady-state kinetic results, which demonstrated that all substrates must be bound before catalysis. Kinetic characterization of a mutant, which releases formyl phosphate into solution, and positional isotope exchange studies also support the assignment of formyl phosphate as a plausible intermediate.

Original languageEnglish (US)
Pages (from-to)6709-6716
Number of pages8
JournalBiochemistry
Volume36
Issue number22
DOIs
Publication statusPublished - Jun 3 1997

    Fingerprint

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this