Closing in on the toxic domain through analysis of a variant Clostridium difficile cytotoxin B.

Abstract:

:Strain 1470 is the standard typing strain for serogroup F of Clostridium difficile containing both toxin genes, toxA-1470 and toxB-1470. A polymerase chain reaction (PCR)-based approach to the sequencing of the total toxB-1470 gene identified an open reading frame (ORF) of 7104 nucleotides. In comparison with the previously sequenced toxB of C. difficile VP10463, the toxB-1470 gene has 16 additional nucleotides, 13 within the 5'-untranslated region and three within the coding region. The M(r) of ToxB-1470 is 269,262, with an isoelectric point (IP) of 4.16. The equivalent values for ToxB are M(r) 269,709 and IP 4.13. In comparison with ToxB, ToxB-1470 differs primarily in the N-terminal region between positions 1 and 868 where 148 amino acids residues are changed. The C-terminal region between residues 869-2367 is highly conserved with only six amino acid alterations. Dot matrix comparison of ToxB-1470 with ToxA and ToxB reveals the highest homology between ToxB-1470 and ToxB. Thus ToxB-1470 did not originate from recombination between ToxA and ToxB. On cultured endothelial cells, from porcine pulmonary artery, purified ToxB-1470 is less potent than ToxB. The cytopathic effects of ToxB-1470 are indistinguishable from those caused by the lethal toxin (LT) of Clostridium sordellii, but are clearly different from the patterns observed after exposure of endothelial cells to ToxA and ToxB of C. difficile (VPI10463) or alpha-toxin (Tcn alpha) of Clostridium novyi. The LT-like action of ToxB-1470 was not due to altered internalization processes, as microinjection and addition to the medium induced identical effects on the cells.(ABSTRACT TRUNCATED AT 250 WORDS)

journal_name

Mol Microbiol

journal_title

Molecular microbiology

authors

von Eichel-Streiber C,Meyer zu Heringdorf D,Habermann E,Sartingen S

doi

10.1111/j.1365-2958.1995.mmi_17020313.x

subject

Has Abstract

pub_date

1995-07-01 00:00:00

pages

313-21

issue

2

eissn

0950-382X

issn

1365-2958

journal_volume

17

pub_type

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