title: Proteomic analysis of the sarcosine-insoluble outer membrane fraction of the bacterial pathogen Bartonella henselae.
abstract: Bartonella henselae is an emerging zoonotic pathogen causing a wide range of disease manifestations in humans. In this study, we report on the analysis of the sarcosine-insoluble outer membrane fraction of B. henselae ATCC 49882 Houston-1 by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1-D SDS-PAGE) and two-dimensional nonequilibrium pH gradient polyacrylamide gel electrophoresis (2-D NEPHGE). Protein species were identified by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) and subsequent database query against the B. henselae genome sequence. Subcellular fractionation, application of the ionic detergent lauryl sarcosine, assessment of trypsin sensitivity, and heat modifiability of surface-exposed proteins represented valuable tools for the analysis of the outer membrane subproteome of B. henselae. 2-D NEPHGE was applied to display and catalogue a substantial number of proteins associated with the B. henselae sarcosine-insoluble outer membrane fraction, resulting in the establishment of a first 2-D reference map of this compartment. Thus, 53 distinct protein species associated with the outer membrane subproteome fraction were identified. This study provides novel insights into the membrane biology and the associated putative virulence factors of this pathogen of increasing medical importance.
authors.affiliation: Division of Molecular Microbiology, Biozentrum of the University of Basel, Basel, Switzerland.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Rhomberg
authors.forename: Olof
authors.initials: O
authors.lastname: Karlberg
authors.forename: Thierry
authors.initials: T
authors.lastname: Mini
authors.forename: Ursula
authors.initials: U
authors.lastname: Zimny-Arndt
authors.forename: Ulrika
authors.initials: U
authors.lastname: Wickenberg
authors.forename: Marlene
authors.initials: M
authors.lastname: Röttgen
authors.forename: Peter R
authors.initials: PR
authors.lastname: Jungblut
authors.forename: Paul
authors.initials: P
authors.lastname: Jenö
authors.forename: Siv G E
authors.initials: SG
authors.lastname: Andersson
authors.forename: Christoph
authors.initials: C
authors.lastname: Dehio
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D018416:Bartonella henselae / Q000235:genetics* / Q000472:pathogenicity*; D002138:Calibration; D030562:Databases, Protein; D003902:Detergents / Q000494:pharmacology; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D016680:Genome, Bacterial; D006863:Hydrogen-Ion Concentration; D007477:Ions; D013058:Mass Spectrometry; D011506:Proteins / Q000737:chemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D012521:Sarcosine / Q000737:chemistry* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013347:Subcellular Fractions / Q000378:metabolism; D014357:Trypsin / Q000494:pharmacology
chemical_list: D001426:Bacterial Proteins; D003902:Detergents; D007477:Ions; D011506:Proteins; D020543:Proteome; D014357:Trypsin; D012521:Sarcosine