Daff Group

Group Members

Davide Papale (PhD student), Andy Welland (PhD student), Sidong Liu (PhD student), Ben Gazur (PhD student):  

NO Synthase

The nitric oxide synthases produce the signalling molecule NO in mammalian systems and are involved in immune response, controlling blood flow and neural development, along with many other proposed functions. They are regulated both transcriptionally and via a series of interdependent mechanisms involving ligand-enzyme binding. These regulation mechanisms are being investigated on the molecular level using a host of techniques, including protein engineering/mutagenesis, steady-state and stopped-flow kinetics, redox potentiomentry and the general tools of protein biochemistry. Conducted in collaboration with Prof. S. K. Chapman (Edinburgh), Prof. G. A. Reid (Edinburgh) and Prof. T. Shimizu (Sendai, Japan). 

Bioelectrochemistry

Quantitative electrochemical analysis of redox enzymes and construction of engineered redox enzymes with enhanced electron transfer properties. Current enzymes under analysis include NO synthase, cytochrome P450 BM3, flavodoxin and other flavoenzymes/cytochromes. 

Cytochrome P450 BM3

A novel, self-sufficient bacterial cytochrome P450 (P450 BM3) being used as a model P450 monooxygenase system. We are using site-directed mutagenesis to modulate the reduction potential of the heme centre, stabilising catalytic intermediates and defining the enzyme’s mechanism in terms of thermodynamic and kinetic parameters. This work should lead to a greater understanding of how oxygen activation occurs in the P450 enzymes.  Conducted in colloboration with Prof. S.K. Chapman (Edinburgh).  

	Address :	School of Chemistry	0131 650 7378
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		West Mains Rd	Simon.Daff@ed.ac.uk
		Edinburgh   EH9 3JJ