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Structural and functional studies on the copper-containing particulate methane monooxygenase from Methylococcus capsulatus (Bath)

Reference	BB/C00194X/1
Principal Investigator / Supervisor	Professor H Dalton
Co-Investigators / Co-Supervisors	Dr Nataliia Myronova
Institution	University of Warwick
Department	Biological Sciences
Funding type	Research
Value (£)	214,713
Status	Completed
Type	Research Grant
Start date	01/08/2004
End date	31/07/2007
Duration	36 months

Abstract

The membrane-associated form of methane monooxygenase has been known for almost thirty years but has eluded resolution and purification into its component forms. We have now achieved purification of the hydroxylase component and have good indications that the electron-donating component is resolvable. We have good analytical and spectroscopic evidence that the active site comprises both copper and ion. This project is aimed at understanding how the active site of the hydroxylase is organised to activate methane and/or oxygen to produce methanol. Spectroscopic techniques that include EPR, ENDOR and possibly Mossbauer and EXAFS will be used to characterise the electronic interactions of metals and ligands at the active site. In addition we have recently, in collaboration with Prof Iwata, managed to obtain good quality crystals of the hydroxylase which will be the basis for out attempt to obtain a 3D structure of the enzyme by x-ray crystallography. Our recent identification of a 32kDa protein, which has an important role in methane activation by the hydroxylase, may be a conformational variant of the structural 47kDa protein of the hydroxylase. We will be using CD and fluorescence spectroscopy to investigate the interplay between the 32 and 47kDa proteins and be asking what role each of these proteins play in the substrate activation process. Electron transfer to the hydroxylase we believe, is mediated by an NDH type 2 enzyme and possibly involves quinols in the process. We will be using both reconstitution studies and proteomics to facilitate the identification of those components of the electron transport chain.

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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