title: Identification and characterization of the Sulfolobus solfataricus P2 proteome.
abstract: Via combined separation approaches, a total of 1399 proteins were identified, representing 47% of the Sulfolobus solfataricus P2 theoretical proteome. This includes 1323 proteins from the soluble fraction, 44 from the insoluble fraction and 32 from the extra-cellular or secreted fraction. We used conventional 2-dimensional gel electrophoresis (2-DE) for the soluble fraction, and shotgun proteomics for all three cell fractions (soluble, insoluble, and secreted). Two gel-based fractionation methods were explored for shotgun proteomics, namely: (i) protein separation utilizing 1-dimensional gel electrophoresis (1-DE) followed by peptide fractionation by iso-electric focusing (IEF), and (ii) protein and peptide fractionation both employing IEF. Results indicate that a 1D-IEF fractionation workflow with three replicate mass spectrometric analyses gave the best overall result for soluble protein identification. A greater than 50% increment in protein identification was achieved with three injections using LC-ESI-MS/MS. Protein and peptide fractionation efficiency; together with the filtration criteria are also discussed.
authors.affiliation: Biological and Environmental Systems Group, Department of Chemical and Process Engineering, University of Sheffield, Mappin Street, Sheffield S1 3JD, United Kingdom.
authors.forename: Poh Kuan
authors.initials: PK
authors.lastname: Chong
authors.forename: Phillip C
authors.initials: PC
authors.lastname: Wright
mesh_terms: D019843:Archaeal Proteins / Q000737:chemistry*; D002850:Chromatography, Gel; D016208:Databases, Factual; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D013058:Mass Spectrometry; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D013347:Subcellular Fractions; D048229:Sulfolobus solfataricus / Q000378:metabolism*; D014357:Trypsin / Q000737:chemistry
chemical_list: D019843:Archaeal Proteins; D010455:Peptides; D011506:Proteins; D020543:Proteome; D014357:Trypsin