Heat‐Stable Enterotoxin Produced by <i>Yersinia enterocolitica</i> Isolated from Patients

Okamoto, Keinosuke; Ichikawa, Hidetaka; Kawamoto, Yoko; Miyama, Akio; Yoshii, Saiji

doi:10.1111/j.1348-0421.1980.tb02844.x

Cited by 23 publications

(16 citation statements)

References 26 publications

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“…Infusion broth (BBL Microbiology Systems, Cockeysville, Md., U.S.A.), nutrient agar (Nissui Seiyaku, Tokyo, Japan), CY medium (14), and minimal medium were used for the cultivation of bacteria. Minimal medium (pH 7.2) contained 8.2 g of Na2HPO4-7H20, 2.7 g of KH2PO4, 1.0 g of (NH4)2SO4, 0.1 g of MgSO4-7H20, 5 mg of Ca(NO3)2, and 0.25 mg of FeSO4.7H20 per liter of distilled water.…”

Section: Methodsmentioning

confidence: 99%

Adherence of Yersinia enterocolitica to Mammalian Epithelial Cell Lines

Okamoto¹,

Inoue

Ichikawa

et al. 1980

Microbiology and Immunology

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Section: Methodsmentioning

confidence: 99%

Adherence of Yersinia enterocolitica to Mammalian Epithelial Cell Lines

Okamoto¹,

Inoue

Ichikawa

et al. 1980

Microbiology and Immunology

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“…Preparation of'the crude toxin Y. enterocolitica strain 23 [23] was grown in CY medium (casamino acids and yeast extract broth supplemented with glucose, pH 8.5) as described in [19]. The culture supernatant of the bacteria was treated with protamine sulfate and separated by centrifugation.…”

Section: Mutrriuls and Upparutusmentioning

confidence: 99%

Isolation, primary structure and synthesis of heat-stable enterotoxin produced by Yersinia enterocolitica

Takao¹,

Tominaga²,

Yoshimura³

et al. 1985

Eur J Biochem

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Five heat-stable enterotoxins were isolated from the culture supernatant of Yersinia enterocolitica and purified to homogeneity by DEAE-Sephacel and high-performance liquid chromatographies. They caused acute fluid accumulation in the intestine of suckling mice. The amino acid sequence of one of the enterotoxins was determined to be Ser-Ser-Asp-Trp-Asp-Cys-Cys-Asp-Val-Cys-Cys-Asn-Pro-Ala-Cys-Ala-Gly-Cys, by Edman degradation of its pyridylethylated derivative and a combination of fast atom bombardment mass spectrometry and carboxypeptidase B digestion. This structure was unambiguously confirmed by chemical synthesis. The other enterotoxins had longer or shorter amino acid sequences at their N termini, but the same sequence at their C termini. The six half-cystine residues formed intramolecular disulfide linkages, as shown by measurement of the molecular masses of the enterotoxins by fast atom bombardment mass spectrometry. The sequence of 13 amino acid residues at the C terminus showed similarity to those of heat-stable enterotoxins isolated from enterotoxigenic Escherichiu c d i [Aimoto, S . et al. (1982) high-performance liquid chromatography (HPLC) and determined the primary structure of one of them [22]. Part of the primary structure of Yersiniu ST determined was similar to the active-site sequence of E. coli ST. These similar sequences were deduced to be responsible for the common biological and immunological properties of E. coli ST and Yersinia ST and to be characteristic of, and conserved in heat-stable enterotoxins produced by, enteric bacteria such as enterotoxigenic E. cwli and Y. mtrrocolitica. Therefore, it seemed important to examine whether the other toxic fractions that we isolated from the culture supernatant of Y . enterocolitica also had a similar primary structure.Here we report purification of toxic fractions from the culture supernatant of Y. enterocolitica by a different chromatographic procedure from that used in previous work [22] and determination of their amino acid sequences. We also describe the chemical synthesis of Yersiniu ST, establishing the amino acid sequence of heat-stable enterotoxin of Y . enterocoliticu. MATERIALS AND METHODS Mutrriuls and upparutusReagent-grade chemicals from several suppliers were used and were purified for analytical experiments before use. Carboxypeptidase B and aminopeptidase M were purchased from Boehringer (Mannheim, FRG) and the Protein Research Foundation (Minoh, Japan) respectively. YMC-ODS ( S -5 ) was obtained from Yamamura Chemical Laboratories Co. Ltd (Kyoto, Japan) and packed into columns (4 x 250 mm, 6 x 150 mm and 8 x 300 mm) in our laboratory for preparative experiments. A microsorb psCls column ( 5 pm,

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“…In this connection the formation of a cointegrate or a composite molecule composed of the Ent plasmid and the R plasmid or its antibiotic-resistance determinant (8) is a significant factor because the existence of the antibiotic-resistance marker may become a strong selective advantage for such a plasmid molecule under the pressure of an antibiotic used for therapeutic as well as prophylactic purposes. These considerations lead to the highly conceivable possibility that within a few decades Ent plasmids will spread to various serotypes of E. coli or even to other enteric bacteria as in fact demonstrated in Salmonella (22), Shigella (28), and Yersinia enterocolitica (16,17,31).…”

Section: Discussionmentioning

confidence: 99%

Conjugal Acquisition and Stable Maintenance of Ent Plasmids in Nontoxigenic Wild‐Type Strains of Escherichia coli

Danbara

Arita

Baba³

et al. 1986

Microbiology and Immunology

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In spite of the ability of the genetic determinants for enterotoxin production to be conjugally transferred, mobilized or transposed, enterotoxigenic Escherichia coli (ETEC) isolated from diarrheal patients is restricted to certain serotypes. Four conjugative enterotoxigenic plasmids (Ent plasmids)encoding either a heatlabile enterotoxin or a heat-stable enterotoxin or both and belonging to one of three incompatibility groups IncFI, IncH1, or IncX, were examined for their transferability to and stability in 157 nonenterotoxigenic Escherichia coli strains belonging to various serotypes and 89 clinical isolates nonenterotoxigenic but belonging to those serotypes in which ETEC from diarrheal patients are usually found. The serotypes of the strains to which Ent plasmids were efficiently transferred and in which they were maintained stably were not always the serotypes in which ETEC had usually been found and vice versa. The frequencies of transfer of four Ent and two R plasmids to each of the 157 recipients were correlated with each other, indicating that the frequency of transfer of the plasmid is not determined by a resident plasmid, if there is one, but by a recipient factor which commonly affects transferability to all donors. These results have led to the conclusion that the reason why only certain serotypes are found among ETEC isolated from diarrheal patients is not the ability of these strains specifically and preferentially to acquire and maintain the Ent plasmids.

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Heat‐Stable Enterotoxin Produced by Yersinia enterocolitica Isolated from Patients

Abstract: Twenty-three strains

Cited by 23 publications

References 26 publications

Adherence of Yersinia enterocolitica to Mammalian Epithelial Cell Lines

Adherence of Yersinia enterocolitica to Mammalian Epithelial Cell Lines

Isolation, primary structure and synthesis of heat-stable enterotoxin produced by Yersinia enterocolitica

Conjugal Acquisition and Stable Maintenance of Ent Plasmids in Nontoxigenic Wild‐Type Strains of Escherichia coli

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