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Molecular analysis of the enzymic properties of human poly (ADP-ribose) polymerase using random and site-directed mutagenesis
Reference
C05217
Principal Investigator / Supervisor
Dr Minnie O'Farrell
Co-Investigators /
Co-Supervisors
Institution
University of Essex
Department
Biological Sciences
Funding type
Research
Value (£)
155,697
Status
Completed
Type
Research Grant
Start date
01/04/1996
End date
01/04/1999
Duration
36 months
Abstract
This study uses mutagenesis of the cDNA to make a detailed molecular analysis of the structure and enzymology of the nuclear enzyme, poly (ADP-ribose) polymerase. The catalytic domain of the enzyme will be subject to random mutagenesis using error-prone PCR and loss of function and gain of function mutants will be isolated. Mutants already isolated following chemical mutagenesis will be further explored using site-directed mutagenesis. All the mutants isolated will be sequenced and characterised by estimating Km, Vmax and the kinetics and structure of polymer product formed. Selected mutants will be taken forward for more detailed analysis in an in vitro assay to explore DNA binding, protein-protein interactions and inter- and intramolecular modifications by the enzyme.
Summary
unavailable
Committee
Closed Committee - Biochemistry & Cell Biology (BCB)
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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