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. 1992 Oct;60(10):4213–4220. doi: 10.1128/iai.60.10.4213-4220.1992

Glycoprotein receptors for a heat-stable enterotoxin (STh) produced by enterotoxigenic Escherichia coli.

T Hirayama 1, A Wada 1, N Iwata 1, S Takasaki 1, Y Shimonishi 1, Y Takeda 1
PMCID: PMC257455  PMID: 1328055

Abstract

Glycoprotein receptors for heat-stable enterotoxin STh of enterotoxigenic Escherichia coli in the rat intestinal cell membrane were identified and characterized. Incubation of rat intestinal cell membranes with radioiodinated N-5-azidonitrobenzoyl-STh[5-19] (125I-ANB-STh[5-19]) followed by photolysis resulted in specific radiolabeling of two distinct proteins with M(r)s of 200,000 (designated STR-200A and STR-200B). STR-200A was found to be composed of two molecules of a protein with an M(r) of 70,000 (70-kDa protein), whereas STR-200B was composed of two different protein molecules with M(r)s of 53,000 (53-kDa protein) and 77,000 (77-kDa protein). These proteins showed no guanylate cyclase activity. The 70-kDa protein was labeled most with 125I-ANB-STh[5-19], suggesting that STR-200A is the main receptor protein in the rat intestinal cell membrane. The carbohydrate moieties of STR-200A and STR-200B were examined by enzymatic deglycosylation. The 70-kDa protein of STR-200A was found to contain N-linked high-mannose-type and/or hybrid-type oligosaccharides, and results suggested that it possesses at least three N glycosylation sites. The 53-kDa protein of STR-200B was found to have an N-linked complex-type oligosaccharide side chain. The deglycosylated 70-kDa protein retained activity for binding to STh, suggesting that the carbohydrate moieties of these receptor proteins are not important for binding with STh.

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