title: Heterogeneity in the molecular species of heat-stable enterotoxin of Vibrio cholerae non-O1 expressed by Escherichia coli carrying the cloned toxin gene.
abstract: The biological activity of the heat-stable enterotoxin of Vibrio cholerae non-O1 (NAG-ST) was found to be predominantly associated with the periplasmic extract (about four-fold higher than the culture supernatant) of a recombinant E. coli (JM109) strain carrying the NAG-ST toxin gene. Four molecular species of NAG-ST, two each from the periplasmic extract and culture supernatant of JM109, were purified. Amino acid sequence analysis of the four NAG-ST peptides isolated by HPLC revealed that they all differed from that of the mature 17-amino acid residue NAG-ST released by V. cholerae non-O1. The M(r)-values of the peptides obtained from the periplasmic extract were 4331 and 2785, while those recovered from the culture supernatant were 3154 and 2785. It thus appears that V. cholerae NAG-ST is synthesized as larger molecules in the recombinant E. coli strain. The differences in sizes of the exported NAG-ST molecule could relate to differences in the enzyme cleavage system between E. coli and V. cholerae.
authors.affiliation: Department of Infectious Diseases Research, National Children's Medical Research Center, Tokyo, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Dohi
authors.forename: H
authors.initials: H
authors.lastname: Kasuga
authors.forename: H
authors.initials: H
authors.lastname: Nakao
authors.forename: A
authors.initials: A
authors.lastname: Ogawa
authors.forename: G B
authors.initials: GB
authors.lastname: Nair
authors.forename: T
authors.initials: T
authors.lastname: Takeda
mesh_terms: D000595:Amino Acid Sequence; D004768:Enterotoxins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics; D006358:Hot Temperature; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000302:isolation & purification; D014734:Vibrio cholerae / Q000472:pathogenicity*
chemical_list: D004768:Enterotoxins; D011994:Recombinant Proteins; C037420:stN protein, Vibrio cholerae