Abstract

Pathogenic Yersinia enterocolitica Recombinants species (Y. enterocolitica, Y. pestis, and Y. pseudotuberculosis) use a virulence plasmid-encoded type three secretion system (TTSS) to inject effector proteins into host cells. Y. enterocolitica YscM1 (LcrQ in Y. pestis and Y. pseudotuberculosis) and its homologue YscM2 are regulatory components of TTSS that are also secreted by this transport apparatus. YscM1 and YscM2 share 57% identity and are believed to be functionally equivalent. We have recombinantly expressed and purified YscM1 and YscM2 in Escherichia coli.

After expression as glutathione S-transferase (GST) fusions, purification to near homogeneity was achieved by glutathione-Sepharose affinity chromatography followed by treatment with PreScission protease to cleave GST and gel filtration on a Superdex 75 column. Such recombinant YscM1 and YscM2 bound efficiently to the specific chaperone SycH, indicating proper folding of the purified proteins. Gel filtration analyses revealed that both YscM1 and YscM2 formed homodimers. YscM1 and YscM2 homodimers could dissociate at high ionic strength, indicating that salt bridges essentially contribute to dimerization. Furthermore, we show that YscM1 and YscM2 are susceptible to thrombin cleavage.

1. Product specifications

Size: 100ug or 1mg

Format: Clear sterile-filtered solution.

Formulation: Y.Enterocolitica (O:9) YopH is supplied in 20 mM HEPES buffer pH-7.6, 250 mM NaCl and 20% glycerol.

Purity: Greater than 80.0% as determined by SDS-PAGE.

Recommended applications: 1. It binds to human IgG, IgM and IgA antibodies.2. Immunoblot test with positive/negative sera panels.

2. Preparation

Expression System: E.Coli

Description

Recombinant Yersinia enterocolitica (O:9) YopH produced in E.coli is a non-glycosylated polypeptide chain having a calculated molecular mass of 52,311 Daltons. Y.Enterocolitica (O:9) YopH is expressed with a 10xHis tag at the N-terminus and purified by proprietary chromatographic techniques.

Stability and Storage

Store at 4°C if the entire vial is to be used within 2 to 4 weeks. Store froze at -20°C for longer periods of time. Avoid multiple cycles of freezing and thawing.