title: FoF1-ATP synthase of Streptomyces fradiae ATCC 19609: structural, biochemical, and functional characterization.
abstract: The patterns of protein phosphorylation in inverted membrane vesicles from the strain Streptomyces fradiae ATCC 19609 were investigated to elucidate the mechanisms of regulation of bacterial membrane bound FoF1-ATP synthase. We found for the first time by two-dimensional gel electrophoresis and mass spectrometry that the β- and b-subunits of the FoF1-ATP synthase complex undergo phosphorylation; 20 proteins with known functions were identified. All eight subunits of FoF1-ATP synthase, i.e. α, β, γ, δ, ε, a, b, and c, were cloned into Escherichia coli and expressed as recombinant proteins. Using a crude preparation of serine/threonine protein kinases, we demonstrated the phosphorylation of recombinant γ-, β-, α- and ε-subunits. The β-subunit was phosphorylated both as a recombinant protein and in vesicles. Differential phosphorylation of membrane-bound and recombinant proteins can be attributed to different pools of protein kinases in each preparation; in addition, certain steps of FoF1-ATP synthase assembly and function might be accompanied by individual phosphorylation patterns. The structure of the operon containing all subunits and regulatory protein I was identified. The phylogenetic similarity of FoF1-ATP synthase from Streptomyces fradiae ATCC 19609 with the respective proteins in saprophytic and pathogenic (including Mycobacterium tuberculosis) bacteria was investigated. Thus, bacterial serine/threonine protein kinases are important for the regulation of FoF1-ATP synthase. From the practical standpoint, our results provide a basis for designing targeted antibacterial drugs.
authors.affiliation: Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, 119991, Russia. valerid@rutenia.ru.
authors.forename: M G
authors.initials: MG
authors.lastname: Alekseeva
authors.forename: T A
authors.initials: TA
authors.lastname: Mironcheva
authors.forename: D A
authors.initials: DA
authors.lastname: Mavletova
authors.forename: S M
authors.initials: SM
authors.lastname: Elizarov
authors.forename: N V
authors.initials: NV
authors.lastname: Zakharevich
authors.forename: V N
authors.initials: VN
authors.lastname: Danilenko
mesh_terms: D025181:ATP Synthetase Complexes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D009876:Operon; D010766:Phosphorylation; D010802:Phylogeny; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013302:Streptomyces / Q000737:chemistry / Q000145:classification / Q000201:enzymology* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D021122:Protein Subunits; D025181:ATP Synthetase Complexes