1887

Abstract

Coat proteins from mature spores of two enterotoxin-positive (Ent) and two enterotoxin-negative (Ent) strains of were solubilized using 50 m-dithiothreitol and 1% sodium dodecyl sulphate at pH 9·7, and alkylated using 110 m-iodoacetamide to prevent aggregation. The coat proteins and type A enterotoxin (CPE) were separated by SDS-PAGE and analysed by Western blotting using anti-CPE antibody. As previously reported, CPE aggregated in the presence of SDS, but no aggregation occurred at concentrations below 15 μg CPE ml. Two CPE-related proteins (34 and 48 kDa) were found in the solubilized spore coat protein of Ent strains while only the 48 kDa CPE-related protein was found in the spore coat fraction of Ent strains. CPE-related proteins comprised 2·7% and 0·8% of the total solubilized coat protein of Ent and Ent strains respectively. CPE-related proteins could be extracted from the spores with 1 % SDS alone. They could also be released by disruption of whole spores, indicating that the CPE-related proteins may be in the spore core or trapped between the core and coat layers. The results suggest that CPE is not a major structural component of the coat fraction of spores.

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1989-11-01
2021-10-16
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