VIBRATIONAL AND DYNAMICAL STUDIES OF ENZYMES

Project: Research project

Project Details

Description

DESCRIPTION: (modified from abstract) The dynamics of enzymatic catalysis
reactions will be studied at the molecular level. Raman and IR difference
spectroscopies will be used for static measurements and kinetic Raman and IR
measurements will be used with submillisecond time resolution, down to 10 ns
resolution in some cases, with capabilities to 50 ps when necessary, for
dynamic measurements. Emphasis will be on two classes of enzymes: the NAD(P)
linked enzymes and the phosphoryl transfer enzymes, both targets for
antibacterial and anti-cancer pharmaceuticals. Particular attention will be
paid to studying dihydrofolate reductase to understand how the pKa of N5 of the
bound dihydrofolate substrate to the E. coli enzyme is raised four units
compared to solution. This will be probed by measurements of mutant proteins of
different dihydrofolate reductase isoenzymes and of a related protein of the
folate family (dihydroneopterin aldolase). Detailed electrostatic calculations
will also be used to understand this system. In addition, the c-Harvey ras p21
protein, which contains the essential GTPase core of G-proteins, and the
Yersinia enzyme, an example of protein-tyrosine phosphatases, will be studied.
The structures of phosphate ground state and the transition state analog
complexes of the native protein and a series of mutants will be studied. The
kinetics of the binding of ligands to proteins and the motions of flexible
loops will be probed by examining ligand binding to lactate dehydrogenase and
other proteins, and loop motions will be examined in lactate dehydrogenase, the
Yersinia enzyme, and other systems.
StatusFinished
Effective start/end date8/1/997/31/00

Funding

  • National Institute of General Medical Sciences

ASJC

  • Catalysis

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