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Mapping physical and biochemical interactions of multicomponent subcellular assemblies

Reference9810651
Principal Investigator / Supervisor Professor David Rice
Co-Investigators /
Co-Supervisors
Dr Iain Ashmore Murray
Institution University of Sheffield
DepartmentMolecular Biology and Biotechnology
Funding typeResearch
Value (£) 95,760
StatusCompleted
TypeResearch Grant
Start date 14/12/1999
End date 14/02/2002
Duration26 months

Abstract

Histidine-tagged proteins can act as catalysts of oxidative protein crosslinking via a reaction which is limited to proteins which form specific intermolecular associations in solution. We propose to use the Saccharomyces cerevisiae spliceosome as a model system to demonstrate proof of principle in respect of the use of His-tagged proteins as tools to map intracellular protein:protein interactions. His-tagged spliceosomal proteins will be incorporated into yeast spliceosomes in vivo and subjected to oxidative cross-linking. Immunological analysis, using antisera raised against putative interacting (and control, non-interacting) splicing-associated proteins - previously identified by genetic (two-hybrid) methods - will be used to characterise the cross-linked products.

Summary

unavailable
Committee Closed Committee - Biomolecular Sciences (BMS)
Research TopicsX – not assigned to a current Research Topic
Research PriorityX – Research Priority information not available
Research Initiative ROPA 1998 (ROPA1998) [1998]
Funding SchemeX – not Funded via a specific Funding Scheme
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