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Comparative membrane proteome analysis of three B orrelia species
Author(s) -
Gesslbauer Bernd,
Poljak Albina,
Handwerker Claudia,
Schüler Wolfgang,
Schwendenwein Daniel,
Weber Corinna,
Lundberg Urban,
Meinke Andreas,
Kungl Andreas J.
Publication year - 2012
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201100211
Subject(s) - proteome , biotinylation , bacterial outer membrane , membrane protein , biology , biochemistry , proteomics , fractionation , trypsin , chemistry , microbiology and biotechnology , chromatography , membrane , escherichia coli , enzyme , gene
The versatility of the surface of B orrelia , the causative agent of L yme borreliosis, is very important in host–pathogen interactions allowing bacteria to survive in ticks and to persist in a mammalian environment. To identify the surface proteome of B orrelia , we have performed a large comparative proteomic analysis on the three most important pathogenic B orrelia species, namely B . burgdorferi (strain B 31), B . afzelii (strain K 78), and B . garinii (strain PB i). Isolation of membrane proteins was performed by using three different approaches: (i) a detergent‐based fractionation of outer membrane proteins; (ii) a trypsin‐based partial shedding of outer cell surface proteins; (iii) biotinylation of membrane proteins and preparation of the biotin‐labelled fraction using streptavidin. Proteins derived from the detergent‐based fractionation were further sub‐fractionated by heparin affinity chromatography since heparin‐like molecules play an important role for microbial entry into human cells. All isolated proteins were analysed using either a gel‐based liquid chromatography ( LC )‐ MS/MS technique or by two‐dimensional (2 D )‐ LC ‐ MS/MS resulting in the identification of 286 unique proteins. Ninety seven of these were found in all three B orrelia species, representing potential targets for a broad coverage vaccine for the prevention of L yme borreliosis caused by the different B orrelia species.