Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 1988 Aug;56(8):1934-41.
doi: 10.1128/iai.56.8.1934-1941.1988.

ADP-ribosyltransferase mutations in the catalytic S-1 subunit of pertussis toxin

Affiliations

ADP-ribosyltransferase mutations in the catalytic S-1 subunit of pertussis toxin

J T Barbieri et al. Infect Immun. 1988 Aug.

Abstract

The ADP-ribosyltransferase activity of pertussis toxin resides within the S-1 subunit of the toxin. Deletion mapping of a recombinant S-1 subunit produced in Escherichia coli showed that amino acids 2 through 180 are required for ADP-ribosylation of Gi protein. Mutants of the S-1 subunit which lacked either amino acids 2 through 22 or amino acids 153 through 180 failed to express enzyme activity, implicating a functional or structural role for these residues in catalysis. The catalytic carboxy-terminal S-1 deletion, C-180, was found to be more soluble than the recombinant S-1 subunit, making it a useful construct for future structure-function studies on enzyme catalysis. Four independent single-amino-acid substitutions which decreased ADP-ribosyltransferase activity were constructed in the recombinant S-1 subunit. Substitution of Asp-11 by Ser, Arg-13 by Leu, or Trp-26 by Ile decreased enzyme activity to below detectable levels (less than 1.0% of that of the recombinant S-1 subunit). The Glu-139-to-Ser substitution reduced ADP-ribosyltransferase activity to 15% of that of the recombinant S-1 subunit. Both the oxidized and reduced forms of the recombinant S-1 subunit and recombinant S-1 subunits containing single-amino-acid substitutions were degraded through identical immunoreactive tryptic peptides, suggesting that the conformations of the mutants are similar to that of the recombinant S-1 subunit. Identification of noncatalytic forms of the S-1 subunit of pertussis toxin which have conserved protein structure is an initial step in the generation of a recombinant noncatalytic form of pertussis toxin which may be tested as a candidate for an acellular vaccine against Bordetella pertussis.

PubMed Disclaimer

Similar articles

Cited by

References

    1. J Biol Chem. 1974 Apr 10;249(7):2088-97 - PubMed
    1. Infect Immun. 1987 Nov;55(11):2546-53 - PubMed
    1. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4424-7 - PubMed
    1. Infect Immun. 1981 Mar;31(3):1223-31 - PubMed
    1. J Biol Chem. 1982 Apr 10;257(7):3739-46 - PubMed

Publication types

LinkOut - more resources