Characterization of a full-length, active-site mutant of diphtheria toxin.

Abstract:

:A full-length recombinant mutant of diphtheria toxin containing serine in place of a crucial active-site glutamate has been purified and characterized. The serine substitution caused a minor structural alteration in the toxin as measured by trypsinolysis. ADP-ribosyltransferase activity and cytotoxicity of the mutant were both decreased by approximately 500-fold. A similar reduction in cytotoxicity was found when the enzymic fragments of both the wild-type and mutant toxins were introduced into the cytosol of fibroblasts by osmotically lysing pinosomes. The mutation did not alter the binding of the toxin to cell surface receptors and had no apparent effect on membrane translocation. The results suggest that the decreased cytotoxicity of the mutant is solely due to the reduced ADP-ribosyltransferase activity.

journal_name

Arch Biochem Biophys

authors

O'Keefe DO

doi

10.1016/0003-9861(92)90626-8

subject

Has Abstract

pub_date

1992-08-01 00:00:00

pages

678-84

issue

2

eissn

0003-9861

issn

1096-0384

pii

0003-9861(92)90626-8

journal_volume

296

pub_type

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