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Transient-state kinetics of hydride transfer between recombinant domains of bacterial proton- translocating transhydrogenase

Reference	B08280
Principal Investigator / Supervisor	Professor Baz Jackson
Co-Investigators / Co-Supervisors
Institution	University of Birmingham
Department	Sch of Biosciences
Funding type	Research
Value (£)	155,464
Status	Completed
Type	Research Grant
Start date	01/10/1998
End date	01/10/2001
Duration	36 months

Abstract

Transhydrogenase couples the transfer of hydride ion equivalents between NAD(H) and NADP(H) to the translocation of protons across a membrane. We have separately isolated and purified recombinant forms of domain I, which binds NAD(H), and domain III, which binds NADP(H), from the transhydrogenase of Rhodospirillum rubrum. In this project we shall investigate the pre-steady-state kinetics of transhydrogenation in complexes of domains I and III by stopped-flow spectrophotometry. The objectives are (a) to determine whether or not hydride transfer between nucleotides is direct, (b) to identify and characterise intermediate states of the proteins during transhydrogenation, (c) to investigate the role of closure of the domain I mobile loop during transhydrogenation, and (d) to localise the H+ -binding and H+ -release steps in catalytic turnover.

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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