Abstract
This review will focus on established and newly emerging strategies for identifying and characterizing enzyme intermediates using a rapid transient kinetic approach. The merits of this methodology as well as the basics of experimental design are described. Several illustrative examples of PEP-utilizing enzymes have been chosen as they all perform unique, novel chemistries involving enzyme intermediates and have proven to be exciting pharmaceutical targets for antibiotics and herbicides. A novel application of this approach using time-resolved electrospray mass spectrometry to detect chemically labile enzyme intermediates is also discussed.
Publication types
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Research Support, U.S. Gov't, P.H.S.
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Review
MeSH terms
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3-Phosphoshikimate 1-Carboxyvinyltransferase
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Aldehyde-Lyases / biosynthesis
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Alkyl and Aryl Transferases / metabolism
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Catalysis
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Enzymes / metabolism*
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Half-Life
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Kinetics
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Mass Spectrometry
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Molecular Structure
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Pentosephosphates / metabolism
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Phosphoenolpyruvate / chemistry*
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Phosphoenolpyruvate / metabolism*
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Spectrometry, Mass, Electrospray Ionization
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
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Substrate Specificity
Substances
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Enzymes
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Pentosephosphates
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arabinose 5-phosphate
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Phosphoenolpyruvate
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Alkyl and Aryl Transferases
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3-Phosphoshikimate 1-Carboxyvinyltransferase
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2-dehydro-3-deoxyphosphooctonate aldolase
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Aldehyde-Lyases