Proteomic analysis of the sarcosine-insoluble outer membrane fraction of the bacterial pathogen Bartonella henselae

• Published in: Proteomics (Weinheim) Vol. 4; no. 10; pp. 3021 - 3033
• Main Authors: Rhomberg, Thomas A., Karlberg, Olof, Mini, Thierry, Zimny-Arndt, Ursula, Wickenberg, Ulrika, Röttgen, Marlene, Jungblut, Peter R., Jenö, Paul, Andersson, Siv G. E., Dehio, Christoph
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01-10-2004; WILEY‐VCH Verlag; Wiley-VCH
• Subjects: Analytical, structural and metabolic biochemistry; Bacterial pathogenesis; Bacterial Proteins - chemistry; Bacteriology; Bartonella henselae; Bartonella henselae - genetics; Bartonella henselae - pathogenicity; Biologi; Biological and medical sciences; Biology; Calibration; Databases, Protein; Detergents - pharmacology; Electrophoresis, Gel, Two-Dimensional; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; Genome, Bacterial; Hydrogen-Ion Concentration; Ions; Mass Spectrometry; Microbiology; Miscellaneous; NATURAL SCIENCES; NATURVETENSKAP; Outer membrane protein; Proteins; Proteins - chemistry; Proteome; Proteomics - methods; Sarcosine - chemistry; Sarcosine - metabolism; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Subcellular Fractions - metabolism; Trypsin - pharmacology; Two-dimensional gel electrophoresis; Bacteria; Bartonellaceae; Bartonella henselae; External membrane; Proteomics; Bacterial pathogenesis; Mass spectrometry; Outer membrane protein; Two-dimensional gel electrophoresis
• ISSN: 1615-9853; 1615-9861; 1615-9861
• DOI: 10.1002/pmic.200400933

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.