WESTERN BLOTTING ANALYSIS OF HEAT-SHOCK PROTEINS OF RICKETTSIALES ANDOTHER EUBACTERIA

Heat shock proteins (Hsp) of four Rickettsia species, three Bartonella species, two Ehrlichia species, Orientia tsutsugamushi and seventeen other eubacterial species were characterized by the enhanced chemilumi nescence Western blotting (WB) technique with antibodies raised agains t recombinant Hsp from Escherichia coli and purified GroES from R. typ hi. Although E. coil DnaK and GroEL have epitopes that are highly cons erved among the homologous proteins found in Rickettsia, Ehrlichia, O. tsutsugamushi, Bartonella and other Proteobacteria, anti-E. coil DnaK and GroEL monoclonal antibodies (Dasch et al. (1990) Ann. N.Y. Acad. Sci. 590, 352-369) recognize less conserved epitopes. In contrast, epi topes on E. coli DnaJ, GrpE and GroES are much less conserved since an ti-E. coil DnaJ, GrpE and GroES polyclonal antibodies did not recogniz e DnaJ, GrpE or GroES homologues in Rickettsia, Bartonella, Orientia, Ehrlichia and Legionella. Polyclonal antiserum prepared against GroES from R. typhi reacted strongly with purified 10 kDa GroES peptide from Rickettsia and Bartonella, and strongly bound to proteins of varying electrophoretic mobility from Wolbachia, Legionella, Proteus and Shige lla flexneri and more weakly to other GroES homologues including that found in E. coil. Consequently, commercially available anti-DnaJ, anti -GrpE and anti-GroES polyclonal antibodies and anti-DnaK monoclonal an tibody raised against their respective recombinant E. coli Hsp are not , suitable for detection and identification of homologues of these pro teins in a wide range of eubacteria. (C) 1998 Federation of European M icrobiological Societies. Published by Elsevier Science B.V. All right s reserved.