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Protein-protein interactions in the colicin DNase- immunity system analysed by protein engineering kinetics and multidimensional NMR

Reference	B04955
Principal Investigator / Supervisor	Professor Colin Kleanthous
Co-Investigators / Co-Supervisors	Professor Richard James, Professor Geoffrey Robert Moore
Institution	University of East Anglia
Department	Biological Sciences
Funding type	Research
Value (£)	282,644
Status	Completed
Type	Research Grant
Start date	01/01/1996
End date	31/07/1999
Duration	43 months

Abstract

A multidisciplinary strategy comprising structure determination by high field NMR, protein engineering and pre-steady state kinetics will be used to explore the specificity and stability of the colicin E9 DNase/Im9 complex. Mutations of Im9 will be engineered based on our current solution structure to probe i, the stability of this very tight complex (Kd = 0.1fM), ii, the electrostatically steered association with the E9 DNase, iii, specificity differences between Im9 and the other members of the DNase-specific immunity proteins and, in particular, what interactions define cognate and non-cognate protein-protein interactions. At the same time, NMR solution structures of Im9 bound to both cognate and non-cognate DNases will be determined. These structures will further guide the protein engineering element of the application.

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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