A direct NMR method for the measurement of competitive kinetic isotope effects

Jefferson Chan, Andrew R. Lewis, Michel Gilbert, Marie France Karwaski, Andrew J. Bennet

Research output: Contribution to journalArticle

Abstract

We present a technique that uses 13 C NMR spectroscopy to measure kinetic isotope effects on the second-order rate constant (k cat /K m) for enzyme-catalyzed reactions. Using only milligram quantities of isotopically labeled substrates, precise competitive KIEs can be determined while following the ongoing reaction directly in a NMR spectrometer. Our results for the Vibrio cholerae sialidaseĝ€"catalyzed hydrolysis of natural substrate analogs support a concerted enzymatic transition state for these reactions.

Original languageEnglish (US)
Pages (from-to)405-407
Number of pages3
JournalNature chemical biology
Volume6
Issue number6
DOIs
StatePublished - Jun 2010

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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