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.