An iTRAQ characterisation of the role of TolC during electron transfer from Shewanella oneidensis MR-1.
Anodophilic bacteria have the ability to generate electricity in microbial fuel cells (MFCs) by extracellular electron transfer to the anode. We investigated the anode-specific responses of Shewanella oneidensis MR-1, an exoelectroactive Gammaproteobacterium, using for the first time iTRAQ and 2D-LC MS/MS driven membrane proteomics to compare protein abundances in S. oneidensis when generating power in MFCs, and growing in a continuous culture. The regulated dataset produced was enriched in membrane proteins. Proteins shown to be more abundant in anaerobic electroactive anodic cells included efflux pump TolC and an uncharacterised tetratricopeptide repeat (TPR) protein, whilst the TonB2 system and associated uncharacterised proteins such as TtpC2 and DUF3450 were more abundant in microaerobic planktonic cells. In order to validate the iTRAQ data, the functional role for TolC was examined using a δTolC knockout mutant of S. oneidensis. Possible roles for the uncharacterised proteins were identified using comparative bioinformatics. We demonstrate that employing an insoluble extracellular electron acceptor requires multiple proteins involved in cell surface properties. All MS and processed data are available via ProteomeXchange with identifier PXD004090.
Biological and Environmental Systems Group, ChELSI Institute, Department of Chemical and Biological Engineering, The University of Sheffield, Sheffield, UK.
Gregory J S
GJ
Fowler
Biological and Environmental Systems Group, ChELSI Institute, Department of Chemical and Biological Engineering, The University of Sheffield, Sheffield, UK.
Ana G
AG
Pereira-Medrano
Biological and Environmental Systems Group, ChELSI Institute, Department of Chemical and Biological Engineering, The University of Sheffield, Sheffield, UK.
Stephen
S
Jaffe
Bristol Robotics Laboratory, Universities of Bristol and of the West of England, , Frenchay Campus, Bristol, UK.
Grzegorz
G
Pasternak
Biological and Environmental Systems Group, ChELSI Institute, Department of Chemical and Biological Engineering, The University of Sheffield, Sheffield, UK.
Trong Khoa
TK
Pham
Bristol Robotics Laboratory, Universities of Bristol and of the West of England, , Frenchay Campus, Bristol, UK.
Pablo
P
Ledezma
Biological and Environmental Systems Group, ChELSI Institute, Department of Chemical and Biological Engineering, The University of Sheffield, Sheffield, UK.
Simon T E
ST
Hall
Bristol Robotics Laboratory, Universities of Bristol and of the West of England, , Frenchay Campus, Bristol, UK.
Ioannis A
IA
Ieropoulos
Biological and Environmental Systems Group, ChELSI Institute, Department of Chemical and Biological Engineering, The University of Sheffield, Sheffield, UK. phillip.wright@newcastle.ac.uk.|Faculty of Science, Agriculture & Engineering, Newcastle University, Devonshire Building, Newcastle upon Tyne, UK. phillip.wright@newcastle.ac.uk.
Phillip C
PC
Wright
D001674:Bioelectric Energy Sources; D018441:Biofilms; D004560:Electricity; D004566:Electrodes; D004579:Electron Transport; D004583:Electrons; D040901:Proteomics / Q000379:methods*; D020592:Shewanella / Q000737:chemistry / Q000235:genetics*; D053719:Tandem Mass Spectrometry
Bioinformatics; MS driven proteomics; Membrane proteins; Microbial fuel cell (MFC) biofilms; Microbiology; Shewanella oneidensis MR-1; iTRAQ and 2D-LC MS