We examined the role in toxicity of histidine-44 of the A subunit of Escherichia coli enterotoxin which is located in the active site cavity close to glutamic acid-112. Although amino acid substitution of histidine-44 usually renders a mutant toxin unstable to trypsin, one mutant, alanine-44 (His44Ala) was found to be stable. His44Ala did not show any agmatine:ADP- ribosyltransferase activity in the presence or absence of recombinant ADP- ribosylation factor. It showed no diarrheal or rabbit skin permeability activity and was a competitor in enterotoxin-ADP-ribosyltransferase assays containing recombinant ADP-ribosylation factor. These results suggest that like glutamic acid-112, histidine-44 plays an essential role in toxicity. A tentative model, which explains NAD+ catalysis and the transfer of the ADP- ribosyl moiety to a target amino acid, is proposed for histidine-44 and glutamic acid-112.
All Science Journal Classification (ASJC) codes
- Molecular Biology