title: Analysis of membrane-protein complexes of the marine sulfate reducer Desulfobacula toluolica Tol2 by 1D blue native-PAGE complexome profiling and 2D blue native-/SDS-PAGE.
abstract: Sulfate-reducing bacteria (SRB) obtain energy from cytoplasmic reduction of sulfate to sulfide involving APS-reductase (AprAB) and dissimilatory sulfite reductase (DsrAB). These enzymes are predicted to obtain electrons from membrane redox complexes, i.e. the quinone-interacting membrane-bound oxidoreductase (QmoABC) and DsrMKJOP complexes. In addition to these conserved complexes, the genomes of SRB encode a large number of other (predicted) membrane redox complexes, the function and actual formation of which is unknown. This study reports the establishment of 1D Blue Native-PAGE complexome profiling and 2D BN-/SDS-PAGE for analysis of the membrane protein complexome of the marine sulfate reducer Desulfobacula toluolica Tol2. Analysis of normalized score profiles of >800 proteins in combination with hierarchical clustering and identification of 2D BN-/SDS-PAGE separated spots demonstrated separation of membrane complexes in their native form, e.g. ATP synthase. In addition to the QmoABC and DsrMKJOP complexes, other complexes were detected that constitute the basic membrane complexome of D. toluolica Tol2, e.g. transport proteins (e.g. sodium/sulfate symporters) or redox complexes involved in Na(+) -based bioenergetics (RnfABCDEG). Notably, size estimation indicates dimer and quadruple formation of the DsrMKJOP complex in vivo. Furthermore, cluster analysis suggests interaction of this complex with a rhodanese-like protein (Tol2_C05230) possibly representing a periplasmic electron transfer partner for DsrMKJOP.
authors.affiliation: General and Molecular Microbiology, Institute for Chemistry and Biology of the Marine Environment (ICBM), Carl von Ossietzky University Oldenburg, Oldenburg, Germany.
authors.forename: Lars
authors.initials: L
authors.lastname: Wöhlbrand
authors.affiliation: General and Molecular Microbiology, Institute for Chemistry and Biology of the Marine Environment (ICBM), Carl von Ossietzky University Oldenburg, Oldenburg, Germany.
authors.forename: Hanna S
authors.initials: HS
authors.lastname: Ruppersberg
authors.affiliation: Mathematical Modelling, Institute for Chemistry and Biology of the Marine Environment (ICBM), Carl von Ossietzky University Oldenburg, Oldenburg, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Feenders
authors.affiliation: Mathematical Modelling, Institute for Chemistry and Biology of the Marine Environment (ICBM), Carl von Ossietzky University Oldenburg, Oldenburg, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Blasius
authors.affiliation: Plant Proteomics, Institute of Plant Genetics, Leibniz University Hannover, Hannover, Germany.
authors.forename: Hans-Peter
authors.initials: HP
authors.lastname: Braun
authors.affiliation: General and Molecular Microbiology, Institute for Chemistry and Biology of the Marine Environment (ICBM), Carl von Ossietzky University Oldenburg, Oldenburg, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Rabus
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis* / Q000737:chemistry*; D020564:Deltaproteobacteria / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry*; D063347:Native Polyacrylamide Gel Electrophoresis / Q000379:methods*
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins
keywords: 2D Blue Native; Complexome profiling; Energy metabolism; Membrane-protein complexes; Microbiology; SDS-PAGE; Sulfate-reducing bacteria