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Redox behaviour of the haem domain of flavocytochrome c(3) from Shewanella frigidimarina probed by NMR

Redox behaviour of the haem domain of flavocytochrome c(3) from Shewanella frigidimarina probed by NMR
Redox behaviour of the haem domain of flavocytochrome c(3) from Shewanella frigidimarina probed by NMR
Flavocytochrome c(3) from Shewanella frigidimarina WO is a tetrahaem periplasmic protein of 64 kDa with fumarate reductase activity. This work reports the first example of NMR techniques applied to the assignment of the thermodynamic order of oxidation of the four individual haems for such large protein, expanding its applicability to a wide range of proteins. NMR data from partially and fully oxidised samples of fcc(3) and a mutated protein with an axial ligand of haem IV replaced by alanine were compared with calculated chemical shifts, allowing the structural assignment of the signals and the unequivocal determination of the order of oxidation of the haems. As oxidation progresses the fcc(3) haem domain is polarised, with haems I and II much more oxidised than haems III and IV, haem IV being the most reduced. Thus, during catalysis as an electron is taken by the flavin adenosine dinucleotide from haem IV, haem III is eager to re-reduce haem IV, allowing the transfer of two electrons to the active site.
shewanella, flavocytochrome, fumarate, respiration, nuclear magnetic resonance, electron transfer protein, multihaemsoluble fumarate reductase, electron-transfer mechanisms, ytochromec(3), desulfovibrio-gigas, vulgaris, ncimb400, identification, cooperativity, spectroscopy, catalyst
0014-5793
185-190
Pessanha, Miguel
7e563109-5e0f-4949-b0cc-9cab7a8e2809
Rothery, Emma L.
522392a8-4d43-45f6-a814-ce80b742d7dc
Louro, Ricardo O.
8014cf88-eb58-4ecd-8e04-8ce90a4e43f6
Turner, David L.
6c4c1ccb-a6f2-47cf-944e-e69269198a29
Miles, Caroline S.
772c7c83-6cd0-4e39-b86c-c2898d7ec125
Reid, GraemeA.
bc46cf4d-a752-4ca3-ab28-df31d36d3551
Chapman, Stephen K.
c8004be1-78bc-406c-8150-2cc282f910fa
Xavier, António V.
77269a6a-27cb-4c75-abf0-7d03ac64c228
Salgueiro, Carlos A.
a9805ac9-e610-42ca-bb85-ef4e288eeffb
Pessanha, Miguel
7e563109-5e0f-4949-b0cc-9cab7a8e2809
Rothery, Emma L.
522392a8-4d43-45f6-a814-ce80b742d7dc
Louro, Ricardo O.
8014cf88-eb58-4ecd-8e04-8ce90a4e43f6
Turner, David L.
6c4c1ccb-a6f2-47cf-944e-e69269198a29
Miles, Caroline S.
772c7c83-6cd0-4e39-b86c-c2898d7ec125
Reid, GraemeA.
bc46cf4d-a752-4ca3-ab28-df31d36d3551
Chapman, Stephen K.
c8004be1-78bc-406c-8150-2cc282f910fa
Xavier, António V.
77269a6a-27cb-4c75-abf0-7d03ac64c228
Salgueiro, Carlos A.
a9805ac9-e610-42ca-bb85-ef4e288eeffb

Pessanha, Miguel, Rothery, Emma L., Louro, Ricardo O., Turner, David L., Miles, Caroline S., Reid, GraemeA., Chapman, Stephen K., Xavier, António V. and Salgueiro, Carlos A. (2004) Redox behaviour of the haem domain of flavocytochrome c(3) from Shewanella frigidimarina probed by NMR. FEBS Letters, 578 (1-2), 185-190. (doi:10.1016/j.febslet.2004.10.098).

Record type: Article

Abstract

Flavocytochrome c(3) from Shewanella frigidimarina WO is a tetrahaem periplasmic protein of 64 kDa with fumarate reductase activity. This work reports the first example of NMR techniques applied to the assignment of the thermodynamic order of oxidation of the four individual haems for such large protein, expanding its applicability to a wide range of proteins. NMR data from partially and fully oxidised samples of fcc(3) and a mutated protein with an axial ligand of haem IV replaced by alanine were compared with calculated chemical shifts, allowing the structural assignment of the signals and the unequivocal determination of the order of oxidation of the haems. As oxidation progresses the fcc(3) haem domain is polarised, with haems I and II much more oxidised than haems III and IV, haem IV being the most reduced. Thus, during catalysis as an electron is taken by the flavin adenosine dinucleotide from haem IV, haem III is eager to re-reduce haem IV, allowing the transfer of two electrons to the active site.

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More information

Published date: 3 December 2004
Keywords: shewanella, flavocytochrome, fumarate, respiration, nuclear magnetic resonance, electron transfer protein, multihaemsoluble fumarate reductase, electron-transfer mechanisms, ytochromec(3), desulfovibrio-gigas, vulgaris, ncimb400, identification, cooperativity, spectroscopy, catalyst

Identifiers

Local EPrints ID: 20304
URI: http://eprints.soton.ac.uk/id/eprint/20304
ISSN: 0014-5793
PURE UUID: 2f99c907-1f90-4022-ae0d-eced4801724a

Catalogue record

Date deposited: 17 Feb 2006
Last modified: 15 Mar 2024 06:23

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Contributors

Author: Miguel Pessanha
Author: Emma L. Rothery
Author: Ricardo O. Louro
Author: David L. Turner
Author: Caroline S. Miles
Author: GraemeA. Reid
Author: Stephen K. Chapman
Author: António V. Xavier
Author: Carlos A. Salgueiro

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