Transition State Analysis of Enzymatic Reactions

Project: Research project

Project Details

Description

The goals of this research project are to characterize the[unreadable]
transition states of biologically significant enzymes of N-ribosyl bond[unreadable]
scission. The transition states are used as atomic templates to develop the[unreadable]
theory and practice of transition state inhibitor design. Transition state[unreadable]
information and transition state analogies are used to investigate the[unreadable]
nature of enzyme-bound transition states. The enzymes of interest include[unreadable]
the protozoan nucleoside N-ribohydrolase isozymes and purine nucleoside[unreadable]
phosphorylase. Protozoan parasites are purine auxotrophs and use three[unreadable]
isozyme of N-ribohydrolases for purine salvage. The enzymes are not found[unreadable]
in mammals. Purine nucleoside phosphorylase is widely distributed and is[unreadable]
essential for normal T-cell function. Transition state structures for these[unreadable]
enzymes have bee determined by kinetic isotope effect measurements using[unreadable]
normal mode semiemperical and ab initio structural analysis. The first[unreadable]
generation of transition state inhibitors have been synthesized and will be[unreadable]
characterized. Experimental approaches will include: 1) laser and infrared[unreadable]
spectroscopic investigation of the bound calcium-water center in the[unreadable]
nucleoside hydrolases; NMR and Raman spectra of bound transition state[unreadable]
inhibitors using isotope-edite difference analysis; and free-electron[unreadable]
induced infrared lasers to study the actual enzymatic transition state; 2)[unreadable]
x-ray crystallography of complexes with substrate and transition state[unreadable]
analogues to define changes in protein structur in empty enzyme, Michaelis[unreadable]
and transition state complexes; 3) development of theory for predicting[unreadable]
binding energies of transition state inhibitors using trained pattern[unreadable]
recognition; synthesis and testing of likely transition state inhibitors.[unreadable]
The most powerful inhibitors will be tested for function in mice. The[unreadable]
results of these studies are expected to give novel information about the[unreadable]
nature of enzymatic transition states, provide new theory for the design of[unreadable]
transition state inhibitors, and to provide powerful transition state[unreadable]
inhibitors for several clinically relevant disorders.[unreadable]
StatusFinished
Effective start/end date8/1/987/31/08

Funding

  • National Institute of General Medical Sciences: $361,299.00
  • National Institute of General Medical Sciences: $501,372.00
  • National Institute of General Medical Sciences
  • National Institute of General Medical Sciences: $500,719.00
  • National Institute of General Medical Sciences: $352,355.00
  • National Institute of General Medical Sciences: $488,369.00
  • National Institute of General Medical Sciences: $343,669.00
  • National Institute of General Medical Sciences: $484,379.00

ASJC

  • Radiation
  • Spectroscopy

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