Award details

Chorismate Synthase

Reference	BBS/E/J/40004037
Principal Investigator / Supervisor	Professor R Thorneley
Co-Investigators / Co-Supervisors	Professor David Lawson, Professor David John Lowe
Institution	John Innes Centre
Department	John Innes Centre Department
Funding type	Research
Value (£)	35,126
Status	Completed
Type	Institute Project
Start date	01/04/1997
End date	31/03/2001
Duration	48 months

Abstract

The rational design of inhibitors for commercial exploitation as antimicrobial agents and/or herbicides is increasingly dependent on structural and mechanistic knowledge, at atomic level resolution, of the target enzyme or receptor. E. coli chorismate synthase is an attractive target because it is the seventh enzyme of the shikimate pathway that leads to essential aromatic amino acid and organic cofactor synthesis in bacteria, fungi and plants. It is not present in mammals. An interdisciplinary team of spectroscopists and kineticists together with a protein crystallographer are elucidating the novel chemistry by which 5- enolpyruvyl shikimate-3-phosphate (EPSP) is converted to chorismate with the participation of a reduced flavin cofactor. Perturbation of the partial reactions that comprise the catalytic cycle by using fluoro-EPSP substrate analogues (derived from fluoro-shikimates which have antimicrobial activity), flavin analogues and isotopically labelled EPSP with transient kinetic, ENDOR (RO 4043) and spectrofluorometric analysis is a feature of the current work on modes of flavin and substrate binding. The enzyme has recently been crystallised and the procedure is being optimised to improve the quality of the X-ray diffraction data for a structure determination. When a structure has been obtained site directed mutagenesis of key active site residues will be initiated to test mechanistic hypotheses. Computer modelling studies will be used to aid the design of inhibitors. Rapid- freeze EPR and solid state NMR are being used to characterise postulated intermediates formed in the catalytic cycle

Summary

unavailable

Committee	Closed Committee - Biomolecular Sciences (BMS)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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