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Engineering haem binding sites in monomeric rop
Reference
E09651
Principal Investigator / Supervisor
Professor Gianfranco Gilardi
Co-Investigators /
Co-Supervisors
Institution
Imperial College London
Department
Biological Sciences
Funding type
Research
Value (£)
184,596
Status
Completed
Type
Research Grant
Start date
01/07/1998
End date
01/07/2001
Duration
36 months
Abstract
This project proposes to engineer haem-binding sites in Rop, a stable and highly expressed helix bundle. The aim is to engineer new biomaterials with the desired redox properties, with the long term objective of creating protein-modified electrodes. Rop is expressed at high (gram / L) levels, it is stable and preliminary experiments have proved its versatility in accommodating structural adjustments. This opens the possibility of inserting the haem prosthetic group during the purification procedure of suitable mutants. These objectives will be achieved by application of site-directed mutagenesis on a monomeric variant of Rop already constructed, expressed and characterised. Unique His and Met pairs will be inserted in the right position to allow for haem binding, already proven possible by preliminary modelling studies. Structural (NMR and EPR) and functional (electrochemistry) characterisation of the resulting molecules will be followed by preliminary immobilisation studies on gold electrodes via unique surface Cys residues.
Summary
unavailable
Committee
Closed Committee - Engineering & Biological Systems (EBS)
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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