The mechanism of the amidases: Mutating the glutamate adjacent to the catalytic triad inactivates the enzyme due to substrate mispositioning

Brandon W. Weber, Serah W. Kimani, Arvind Varsani, Donald A. Cowan, Roger Hunter, Gerhard A. Venter, James C. Gumbart, B. Trevor Sewell

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

Background:A cysteine, a glutamic acid, and a lysine are the well known amidase catalytic residues. Results:Mutating the neighboring, structurally conserved Glu-142 inactivates the enzyme, but the active site cysteine still reacts with acrylamide via its double bond. Conclusion:Glu-142 positions the amide for productive nucleophilic attack by the cysteine. Significance:An intact catalytic tetrad is required for amidase activity.

Original languageEnglish (US)
Pages (from-to)28514-28523
Number of pages10
JournalJournal of Biological Chemistry
Volume288
Issue number40
DOIs
StatePublished - Oct 4 2013
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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