A functional description of CymA, an electron-transfer hub supporting anaerobic respiratory flexibility in Shewanella

Authors

Sophie J. Marritt, University of East Anglia
Thomas G. Lowe, University of East Anglia
Jordan Bye, University of East Anglia
Duncan G. G. Mcmillan, University of Leeds
Liang Shi, Pacific Northwest National LaboratoryFollow
James K. Fredrickson, Pacific Northwest National LaboratoryFollow
John M. Zachara, Pacific Northwest National LaboratoryFollow
David J. Richardson, University of East AngliaFollow
Myles R. Cheesman, University of East Anglia
Lars J. C. Jeuken, University of Leeds
Julea N. Butt, University of East Anglia

Date of this Version

2012

Citation

Biochem. J. (2012) 444, 465–474; doi:10.1042/BJ20120197

Abstract

CymA (tetrahaem cytochrome c) is a member of the NapC/NirT family of quinol dehydrogenases. Essential for the anaerobic respiratory flexibility of shewanellae, CymA transfers electrons from menaquinol to various dedicated systems for the reduction of terminal electron acceptors including fumarate and insoluble minerals of Fe(III). Spectroscopic characterization of CymA from Shewanella oneidensis strain MR-1 identifies three lowspin His/His co-ordinated c-haems and a single high-spin chaem with His/H₂O co-ordination lying adjacent to the quinolbinding site. At pH 7, binding of the menaquinol analogue, 2-heptyl-4-hydroxyquinoline-N-oxide, does not alter the midpoint potentials of the high-spin (approximately −240 mV) and low-spin (approximately −110, −190 and −265 mV) haems that appear biased to transfer electrons from the high- to lowspin centres following quinol oxidation. CymA is reduced with menadiol (E_m= −80 mV) in the presence ofNADH(E_m= −320 mV) and an NADH–menadione (2-methyl-1,4-naphthoquinone) oxidoreductase, but not by menadiol alone. In cytoplasmic membranes reduction of CymA may then require the thermodynamic driving force from NADH, formate or H₂ oxidation as the redox poise of the menaquinol pool in isolation is insufficient. Spectroscopic studies suggest that CymA requires a non-haem co-factor for quinol oxidation and that the reduced enzyme forms a 1:1 complex with its redox partner Fcc₃ (flavocytochrome c₃ fumarate reductase). The implications for CymA supporting the respiratory flexibility of shewanellae are discussed.