title: Shotgun proteomics of the haloarchaeon Haloferax volcanii.
abstract: Haloferax volcanii, an extreme halophile originally isolated from the Dead Sea, is used worldwide as a model organism for furthering our understanding of archaeal cell physiology. In this study, a combination of approaches was used to identify a total of 1296 proteins, representing 32% of the theoretical proteome of this haloarchaeon. This included separation of (phospho)proteins/peptides by 2-dimensional gel electrophoresis (2-D), immobilized metal affinity chromatography (IMAC), metal oxide affinity chromatography (MOAC), and Multidimensional Protein Identification Technology (MudPIT) including strong cation exchange (SCX) chromatography coupled with reversed phase (RP) HPLC. Proteins were identified by tandem mass spectrometry (MS/MS) using nanoelectrospray ionization hybrid quadrupole time-of-flight (QSTAR XL Hybrid LC/MS/MS System) and quadrupole ion trap (Thermo LCQ Deca). Results indicate that a SCX RP HPLC fractionation coupled with MS/MS provides the best high-throughput workflow for overall protein identification.
authors.affiliation: Department of Microbiology and Cell Science, University of Florida, Gainesville, Florida 32611-0700, USA.
authors.forename: P Aaron
authors.initials: PA
authors.lastname: Kirkland
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Humbard
authors.forename: Charles J
authors.initials: CJ
authors.lastname: Daniels
authors.forename: Julie A
authors.initials: JA
authors.lastname: Maupin-Furlow
mesh_terms: D019843:Archaeal Proteins / Q000032:analysis* / Q000302:isolation & purification*; D019622:Haloferax volcanii / Q000737:chemistry*; D008954:Models, Biological; D010455:Peptides / Q000032:analysis; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D014357:Trypsin / Q000494:pharmacology
chemical_list: D019843:Archaeal Proteins; D010455:Peptides; D020543:Proteome; D014357:Trypsin