Proteome analysis of Myxococcus xanthus by off-line two-dimensional chromatographic separation using monolithic poly-(styrene-divinylbenzene) columns combined with ion-trap tandem mass spectrometry.
Myxobacteria are potent producers of secondary metabolites exhibiting diverse biological activities and pharmacological potential. The proteome of Myxococcus xanthus DK1622 was characterized by two-dimensional chromatographic separation of tryptic peptides from a lysate followed by tandem mass spectrometric identification. The high degree of orthogonality of the separation system employing polymer-based strong cation-exchange and monolithic reversed-phase stationary phases was clearly demonstrated. Upon automated database searching, 1312 unique peptides were identified, which were associated with 631 unique proteins. High-molecular polyketide synthetases and nonribosomal peptide synthetases, known to be involved in the biosynthesis of various secondary metabolites, were readily detected. Besides the identification of gene products associated with the production of known secondary metabolites, proteins could also be identified for six gene clusters, for which no biosynthetic product has been known so far.
Schley
Christian
C
Department of Chemistry, Instrumental Analysis and Bioanalysis, Saarland University, 66123 Saarbrücken, Germany.
Altmeyer
Matthias O
MO
Swart
Remco
R
Müller
Rolf
R
Huber
Christian G
CG
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics; D002852:Chromatography, Ion Exchange / Q000379:methods*; D005798:Genes, Bacterial; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D016941:Myxococcus xanthus / Q000378:metabolism*; D010446:Peptide Fragments / Q000032:analysis; D010453:Peptide Synthases / Q000032:analysis; D048630:Polyketide Synthases / Q000032:analysis; D011137:Polystyrenes / Q000737:chemistry; D020543:Proteome / Q000032:analysis*; D012333:RNA, Messenger / Q000032:analysis
D001426:Bacterial Proteins; D010446:Peptide Fragments; D011137:Polystyrenes; D020543:Proteome; D012333:RNA, Messenger; D048630:Polyketide Synthases; C003771:Amberlite XAD-2 resin; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase