title: Towards a phosphoproteome map of Corynebacterium glutamicum.
abstract: In this study, the phosphoproteome of Corynebacterium glutamicum, an industrially important soil bacterium of the Corynebacterium/Mycobacterium/Nocardia (CMN) group of Gram-positive bacteria, was investigated by two different detection methods: first, by in vivo radio-labeling using [(33)P]-phosphoric acid with subsequent autoradiography and second, by immunostaining with phosphoamino acid-specific monoclonal antibodies. After two-dimensional gel electrophoresis (2-DE), around 60 [(33)P]-labeled protein spots were visualized and around 90 antibody-decorated protein spots detected; 31 of the protein spots were detected with both methods. By peptide mass fingerprinting, 41 different proteins were identified, namely 5-enolpyruvylshikimate 3-phosphate synthase, aconitase, acyl-CoA carboxylase, acyl-CoA synthetase, ATP (synthase alpha- and beta-chain), carbamoyl-phosphate synthase, citrate synthase, cysteine synthase, DnaK, the elongation factors G, P, Ts and Tu, enolase, fructose bisphosphate aldolase, fumarase, Gap dehydrogenase, glutamine synthetase I, glycine hydroxymethyltransferase, GroEL2, GTPase, heat-inducible transcriptional repressor DnaJ2, inorganic pyrophosphatase, isocitrate dehydrogenase, ketol-acid reductoisomerase, lactate dehydrogenase, leucine-tRNA ligase, lipoamide dehydrogenase, methionine synthase, O-acetylhomoserine sulfhydrylase, pyruvate carboxylase, pyruvate kinase, pyruvate oxidase, ribosomal protein S1, RNA polymerase (beta-subunit), succinyl-CoA:CoA transferase, transketolase and UDP-N-acetylmuramoyl-L-alanine ligase, besides a hypothetical 35k protein and a hypothetical glucose kinase. Both detection techniques were used to create a phosphoproteome map. Additionally, the influence of nitrogen deprivation on the phosphoproteome of C. glutamicum was investigated.
authors.affiliation: Institut für Biochemie, Universität zu Köln, Zülpicher Strasse 47, D-50674 Köln, Germany.
authors.forename: Anne K
authors.initials: AK
authors.lastname: Bendt
authors.forename: Andreas
authors.initials: A
authors.lastname: Burkovski
authors.forename: Steffen
authors.initials: S
authors.lastname: Schaffer
authors.forename: Michael
authors.initials: M
authors.lastname: Bott
authors.forename: Mike
authors.initials: M
authors.lastname: Farwick
authors.forename: Thomas
authors.initials: T
authors.lastname: Hermann
mesh_terms: D003352:Corynebacterium / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D010750:Phosphoproteins / Q000378:metabolism*; D020543:Proteome
chemical_list: D010750:Phosphoproteins; D020543:Proteome