Evidence that the A2 fragment of Shiga-like toxin type I is required for holotoxin integrity

P. R. Austin, P. E. Jablonski, G. A. Bohach, A. Dunker, C. J. Hovde

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Escherichia coli Shiga-like toxin type I (SLT-I) is a potent cytotoxin consisting of an enzymatically active A subunit and a pentameric B subunit that mediates toxin binding to susceptible eukaryotic cells. Evidence that the carboxy-terminal 38 amino acids of the A subunit are involved in holotoxin 1A:5B association is presented. We compared the ability of purified recombinant SLT-I B subunit (Slt-IB) to combine in vitro with purified recombinant SLT-I A subunit (Slt-IA; full-length subunit A includes amino acids 1 to 293) and its ability to combine with purified recombinant SLT-I A1 subunit (Slt-IA1; truncated subunit A includes amino acids 1 to 255). Each mixture was analyzed for biological and physical evidence of toxin assembly. Although Slt-IA successfully combined with Slt-IB to form a molecular species similar to holotoxin that was detectable by nondenaturing polyacrylamide gel electrophoresis and immunoblotting and yielded a molecule which was cytotoxic to cultured Vero cells, Slt-IA1 did not have this ability. Slt-IA1 was 36-fold more active than Slt-IA in an in vitro protein synthesis inhibition assay. These findings suggest that the Slt-IA2 fragment is crucial for formation of SLT holotoxin and stabilizes the interaction between the A and B subunits.

Original languageEnglish (US)
Pages (from-to)1768-1775
Number of pages8
JournalInfection and Immunity
Volume62
Issue number5
StatePublished - 1994
Externally publishedYes

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Shiga Toxin 1
varespladib methyl
Amino Acids
Vero Cells
Cytotoxins
Eukaryotic Cells
Immunoblotting
Polyacrylamide Gel Electrophoresis
Cultured Cells
stichoposide
Escherichia coli
Proteins

ASJC Scopus subject areas

  • Immunology

Cite this

Austin, P. R., Jablonski, P. E., Bohach, G. A., Dunker, A., & Hovde, C. J. (1994). Evidence that the A2 fragment of Shiga-like toxin type I is required for holotoxin integrity. Infection and Immunity, 62(5), 1768-1775.

Evidence that the A2 fragment of Shiga-like toxin type I is required for holotoxin integrity. / Austin, P. R.; Jablonski, P. E.; Bohach, G. A.; Dunker, A.; Hovde, C. J.

In: Infection and Immunity, Vol. 62, No. 5, 1994, p. 1768-1775.

Research output: Contribution to journalArticle

Austin, PR, Jablonski, PE, Bohach, GA, Dunker, A & Hovde, CJ 1994, 'Evidence that the A2 fragment of Shiga-like toxin type I is required for holotoxin integrity', Infection and Immunity, vol. 62, no. 5, pp. 1768-1775.
Austin, P. R. ; Jablonski, P. E. ; Bohach, G. A. ; Dunker, A. ; Hovde, C. J. / Evidence that the A2 fragment of Shiga-like toxin type I is required for holotoxin integrity. In: Infection and Immunity. 1994 ; Vol. 62, No. 5. pp. 1768-1775.
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