Recognition by peptide mapping of three different structural groups of outer membrane protein YOP-1 of Yersinia enterocolitica and Yersinia pseudotuberculosis

Abstract

The structural relation of YOP-1 of "european" and "american" Yersinia enterocolitica serotypes O:3, O:9, O:5,27, and O:8 and O:20, respectively, and Y. pseudotuberculosis serotypes I, II, and III was compared by sodium dodecyl sulfate polyacrylamide gel electrophoresis and peptide mapping using Staphylococcus aureus protease V8. Apparent molecular weights of YOP-1 ranged from 206,000 (O:3) to approx. 180,000 (O:8). According to their respective peptide maps YOP-1 of the "european" and "american" Y. enterocolitica serotypes and Y. pseudotuberculosis serotypes could be assigned to three different groups. Evaluation of several isolates of Y. enterocolitica serotypes O:3, O:9, and O:8 by peptide mapping indicated that YOP-1 is conserved within a serotype. However, one serotype O:8 isolate differed from the consensus peptide pattern of the other serotype O:8 and O:20 isolates. The similarity of the peptide patterns of Yersinia serotypes which predominate in certain geographical locations, i.e., "european" and "american" Y. enterocolitica serotypes, suggest common evolution of YOP-1 of these serotypes independent of the evolution of the other serotypes.

Characterization of different oligomeric species of the Yersinia enterocolitica outer membrane protein YadA

“…4). This atypical behavior may in part explain the wide range of reported apparent M r of YadA (Cornelis et al 1987;Heesemann and Grtiter 1987;Mack et al 1991;Skurnik et al 1984;Zaleska et al 1985).…”

“…6). Reported apparent M r of YadA varied considerably ranging from 160,000 to 240,000 in SDS-PAGE (Cornelis et al 1987;Heesemann and Griiter 1987;Mack et al 1991;Skurnik et al 1984;Zaleska et al 1985). The YadA molecule was reported to dissociate into identical subunits of M r 50,000 on heating in the presence of 10 M urea and 2-mercaptoethanol in gel sample buffer (Skurnik et al 1984;Zaleska et al 1985) and was proposed to be stabilized by disulfide bonds which were highly resistant to reduction due to the conformation of YadA (Skurnik et al 1984).…”

“…A molecular weight of 44,439 is predicted from the amino acid sequence of YadA of Y. enterocolitica serotype O : 3 . However, YadA exhibits an apparent M r ranging from 160,000 to 240,000 in SDS-PAGE depending on the individual Yersinia strain examined (Cornelis et al 1987;Heesemann and Grtiter 1987;Mack et al 1991;Skurnik et al 1984;Zaleska et al 1985). Therefore, YadA was proposed to form a tetramer .…”

“…Myosin (M r 212,000), ~-macroglobulin (M r 170,000), ]%galactosidase (M r 116,000), phosphorylase b (M r 94,000), transferrin (M r 76,000), albumin (M T 67,000), glutamic dehydrogenase (M r 53,000), ovalbumin (M r 43,000), carbonic anhydrase (M r 30,000), and trypsin inhibitor (M r 20,000), were obtained from Pharmacia and were run in parallel as molecular weight markers. In situ peptide mapping of YadA species using Staphylococcus aureus protease V8 was performed as described previously (Cleveland et al 1977;Mack et al 1991). Proteins were stained with Coomassie blue or were detected by silver staining (Morrissey 1981).…”

Experimental Yersinia enterocolitica infection in rats: analysis of the immune response to plasmid-encoded antigens of arthritis-susceptible Lewis rats and arthritis-resistent Fischer rats

“…Partial digestion of YadA by V8 protease was performed as described recently by Mack et al (1991). YadA fragments are superior for antibody detection than the intact YadA antigen because of reduction of unspecific binding of immunoglobulin.…”