A single amino acid substitution in the A subunit of Escherichia coli enterotoxin results in a loss of its toxic activity.
A plasmid encoding a mutant gene of heat-labile enterotoxin (LT), produced by enterotoxigenic Escherichia coli, was induced by treatment of plasmid EWD 299 with hydroxylamine. A mutant strain of E. coli HB 101 carrying the mutant plasmid pTUH 6A produced a low toxic LT analogue (mutant LT), which was cross-reactive with anti-LT antibody. The mutant LT activity was less than 0.15 and 0.006% of the normal LT in the rabbit ileal loop test and in the rabbit skin permeability test, respectively. The amino acid composition of the mutant LT-B subunit was the same as that of the normal B subunit. Though the A2 fragment of the mutant LT was identical to normal LT by DNA analysis, the A1 fragment of the mutant LT differed from the normal A1 fragment in one amino acid at position 112; namely it had lysine instead of glutamic acid from the N terminus. These data suggest that glutamic acid at position 112 from the N terminus of the A1 fragment is important for the A subunit to express its biological activity.
Tsuji
T
T
Department of Microbiology, Fujita Health University School of Medicine, Aichi, Japan.
Inoue
T
T
Miyama
A
A
Okamoto
K
K
Honda
T
T
Miwatani
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics* / Q000302:isolation & purification / Q000633:toxicity; D001483:Base Sequence; D002199:Capillary Permeability / Q000187:drug effects; D004768:Enterotoxins / Q000235:genetics* / Q000302:isolation & purification / Q000633:toxicity; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011817:Rabbits; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000633:toxicity; D012689:Sequence Homology, Nucleic Acid; D012867:Skin / Q000098:blood supply
D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D046911:Macromolecular Substances; D011994:Recombinant Proteins; C027538:heat-labile enterotoxin, E coli