Reversion of recombinant toxoids: mutations in diphtheria toxin that partially compensate for active-site deletions.

Abstract:

:Deleting an important active-site residue of diphtheria toxin, glutamic acid-148, reduces the toxin's ADP-ribosyltransferase activity by a factor of greater than 10(4). We considered using this mutation to construct a recombinant toxoid for expression by live attenuated vaccines and explored second-site mutations that might cause reversion. Activity was partially restored by substituting glutamic acid for valine-147 or by extending the deletion by five residues toward the NH2 terminus, thereby placing glutamic acid-142 immediately adjacent to tyrosine-149. In both mutants the indicated glutamic acid may occupy a spatial locus similar to that of glutamic acid-148 in the unmutated protein. Simply deleting a crucial residue does not, therefore, provide confidence that a second-site mutation could not readily restore activity to a toxoid.

authors

Killeen KP,Escuyer V,Mekalanos JJ,Collier RJ

doi

10.1073/pnas.89.13.6207

keywords:

subject

Has Abstract

pub_date

1992-07-01 00:00:00

pages

6207-9

issue

13

eissn

0027-8424

issn

1091-6490

journal_volume

89

pub_type

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