Detection of novel enzyme intermediates in PEP-utilizing enzymes.
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.
Anderson
Karen S
KS
Department of Pharmacology, SHM B350B, Yale University School of Medicine, 333 Cedar Street New Haven, CT 06520, USA. karen.anderson@yale.edu
D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D000446:Aldehyde-Lyases / Q000096:biosynthesis; D019883:Alkyl and Aryl Transferases / Q000378:metabolism; D002384:Catalysis; D004798:Enzymes / Q000378:metabolism*; D006207:Half-Life; D007700:Kinetics; D013058:Mass Spectrometry; D015394:Molecular Structure; D010428:Pentosephosphates / Q000378:metabolism; D010728:Phosphoenolpyruvate / Q000737:chemistry* / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity
D004798:Enzymes; D010428:Pentosephosphates; C028055:arabinose 5-phosphate; D010728:Phosphoenolpyruvate; D019883:Alkyl and Aryl Transferases; D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; C020011:2-dehydro-3-deoxyphosphooctonate aldolase; D000446:Aldehyde-Lyases