Purification and some properties of a Vero toxin from Escherichia coli O157:H7 that is immunologically unrelated to Shiga toxin

Yutsudo, Takashi; Nakabayashi, Naomi; Hirayama, Toshiya; Takeda, Yoshifumi

doi:10.1016/0882-4010(87)90034-9

Cited by 134 publications

(101 citation statements)

References 26 publications

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“…VTl is biologically, immunochemically and physicochemically identical to Shiga toxin (Noda et al, 1987;Padhye et al, 1989b). Although the biological activity of VT2 is similar to that of VT1, physicochemical analysis has indicated differences in their isoelectric points (Padhye et al, 1987(Padhye et al, , 1989bYutsudo et al, 1987;Downes et al, 1988) and differences in the mol. wts Of their A and subunits-These together Fig.…”

Section: Discussionmentioning

confidence: 99%

“…Both VT1 and VT2 consist of one subunit A and multiple copies of subunit B (Noda et al, 1987;Padhye et al, 1987Padhye et al, , 1988Padhye et al, and 1989bYutsudo et al, 1987). Subunit B is presumed to bind specifically to the Vero cell-surface receptor and mediate uptake of the toxins, whereas subunit A causes inhibition of protein synthesis (Igarashi et al, 1987 ;Ogasawara et al, 1988) and subsequent cell death.…”

Section: Introductionmentioning

confidence: 99%

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Production and characterisation of monoclonal antibodies to Verotoxins 1 and 2 from Escherichia coli of serotype O157:H7

Padhye

Zhao²,

Doyle³

1989

Journal of Medical Microbiology

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Summary. Fourteen hybridoma cell lines were isolated that produced monoclonal antibodies (MAbs) to purified Verotoxins 1 and 2 (VT1, VT2) of Escherichia coli of serotype 0 1 57 : H7. Of these MAbs, eight were obtained by immunisation of BALB/c mice with purified VT1, and six were obtained from BALB/c mice immunised with purified VT2. With the exception of MAb 1C5, with a heavy chain of IgGZb class, antibodies produced from mice immunised with heat-treated toxin were of IgM class. MAbs produced from mice immunised with heat-treated VT1 or VT2 reacted with both verotoxins in ELISA, and Western-blot analysis revealed that they reacted with subunit A and the Al fragment of nicked subunit A of both toxins, but not with subunit B; furthermore, none of them neutralised Vero cytotoxicity or mouse lethality of either toxin. In contrast, MAbs produced from mice immunised with heat-treated and formalin-treated VTl reacted in Western blots with subunits A and B of VT1 and subunit A, but not subunit B, of VT2, reacted in ELISA with VT1 only, and neutralised Vero cytotoxicity and mouse lethality of VT1 but not of VT2. Results indicate the existence of a common epitope on subunit A of VT1 and VT2 that is not responsible for the biological activity of these toxins, and that subunit B is essential for the biological activity of VT1. MAbs capable of reacting with both verotoxins from E. coli of serotype 0157:H7 may be useful reagents for screening bacterial isolates capable of producing one or both of these toxins.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Production and characterisation of monoclonal antibodies to Verotoxins 1 and 2 from Escherichia coli of serotype O157:H7

Padhye

Zhao²,

Doyle³

1989

Journal of Medical Microbiology

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“…Each recombinant E. coli was cultured in Luria-Bertani (LB) broth with 100 g/ml ampicillin (for Stx1) or Mueller-Hinton broth with 5 g/ml trimethoprim (for Stx2) at 37°C for 2 days with vigorous shaking. Stx1 and Stx2 were purified by DEAE-Sephacel (for Stx1) or DEAE-Sepharose (for Stx2) column chromatography, PBE94 chromatofocusing chromatography (Amersham Pharmacia Biotech), and TSK-gel G-2000 SW (Tohso) as described previously (13,14).…”

Section: Methodsmentioning

confidence: 99%

Serum Amyloid P Component Is the Shiga Toxin 2-neutralizing Factor in Human Blood

Kimura

Tani

Matsumoto

et al. 2001

Journal of Biological Chemistry

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Because analysis of this factor is important to understanding the pathology induced by Shiga toxin-producing Escherichia coli, we purified this factor from human plasma and identified it. Purification was carried out by serially subjecting human plasma to Con A-Sepharose, DEAE-Sepharose, hydroxyapatite, and gel-filtration high performance liquid chromatography (HPLC), using Stx2-neutralizing activity as the indicator. The gel-filtration HPLC fraction yielded a single band on SDSpolyacrylamide gel electrophoresis. Twenty N-terminal amino acid residues of this fraction were analyzed and found to correspond perfectly to human serum amyloid P component (HuSAP). Because commercially available HuSAP also showed Stx2 binding and neutralizing activity, we identified this factor as HuSAP.

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“…Toxin structure, function and mode of action Both VTl and VT2 have been purified to homogeneity and, like Shiga toxin, both VTs are made up of A and B subunits (Yutsudo et al, 1987;reviewed by O'Brien and Holmes, 1987). Based on the translated nucleotide sequences the mol.…”

Section: Properties Of E Coli Strains Producing Vtmentioning

confidence: 99%

Vero cytotoxin-producing strains of Escherichia coli

Smith

Scotland

1988

Journal of Medical Microbiology

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Purification and some properties of a Vero toxin from Escherichia coli O157:H7 that is immunologically unrelated to Shiga toxin

Cited by 134 publications

References 26 publications

Production and characterisation of monoclonal antibodies to Verotoxins 1 and 2 from Escherichia coli of serotype O157:H7

Production and characterisation of monoclonal antibodies to Verotoxins 1 and 2 from Escherichia coli of serotype O157:H7

Serum Amyloid P Component Is the Shiga Toxin 2-neutralizing Factor in Human Blood

Vero cytotoxin-producing strains of Escherichia coli

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