Multidimensional proteomic analysis of the soluble subproteome of the emerging nosocomial pathogen Ochrobactrum anthropi.
We report the first large-scale gel-free proteomic analysis of the soluble subproteome of the emerging pathogen Ochrobactrum anthropi. Utilizing our robust offline multidimensional protein identification protocol, a total of 57 280 peptides were initially identified utilizing automated MS/MS analysis software. We describe our investigation of the heuristic protein validation tool PROVALT and demonstrate its ability to increase the speed and accuracy of the curation process of large-scale proteomic datasets. PROVALT reduced our peptide list to 8517 identified peptides and further manual curation of these peptides led to a final list of 984 uniquely identified peptides that resulted in the positive identification of 249 proteins. These identified proteins were functionally classified and physiochemically characterized. A variety of typical "housekeeping" functions identified within the proteome included nucleic acid, amino and fatty acid anabolism and catabolism, glycolysis, TCA cycle, and pyruvate and selenoamino acid metabolism. In addition, a number of potential virulence factors of relevance to both plant and human disease were identified.
Graham
Robert Leslie James
RL
School of Biomedical Sciences, University of Ulster, Coleraine, County Londonderry, BT52 1SA, United Kingdom. rl.graham@ulster.ac.uk
Pollock
Catherine E
CE
O'Loughlin
S Naomi
SN
Ternan
Nigel G
NG
Weatherly
D Brent
DB
Jackson
Philip J
PJ
Tarleton
Rick L
RL
McMullan
Geoff
G
D000595:Amino Acid Sequence; D001331:Automation; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D020621:Ochrobactrum anthropi / Q000737:chemistry* / Q000235:genetics / Q000472:pathogenicity; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D020543:Proteome; D040901:Proteomics / Q000379:methods
D001426:Bacterial Proteins; D010446:Peptide Fragments; D020543:Proteome