DOI: 10.1126/science.109563. Dominant-Negative Mutants of a Toxin Subunit: An Approach to Therapy of Anthrax

Dominant-Negative Mutants of a Toxin Subunit: An Approach to Therapy of Anthrax

10.1126/science.109563

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

The protective antigen moiety of anthrax toxin translocates the toxin's enzymic moieties to the cytosol of mammalian cells by a mechanism that depends on its ability to heptamerize and insert into membranes. We identified dominant-negative mutants of protective antigen that co-assemble with the wild-type protein and block its ability to translocate the enzymic moieties across membranes. These mutants strongly inhibited toxin action in cell culture and in an animal intoxication model, suggesting that they could be useful in therapy of anthrax.

Bibliography

Sellman, B. R., Mourez, M., & John Collier, R. (2001). Dominant-Negative Mutants of a Toxin Subunit: An Approach to Therapy of Anthrax. Science, 292(5517), 695â697.

Authors 3

Bret R. Sellman (first)
Michael Mourez (additional)
R. John Collier (additional)

References 31 Referenced 154

R. Rapuoli M. Pizza in Cellular Microbiology P. Cossart P. Boquet S. Normark R. Rappuoli Eds. (American Society for Microbiology Press Washington DC 2000) pp. 193–220.
Vinion-Dubiel A. D., et al., J. Biol. Chem. 274, 37736 (1999). (10.1074/jbc.274.53.37736) / J. Biol. Chem. by Vinion-Dubiel A. D. (1999)
10.1056/NEJM199909093411107
10.1126/science.280.5364.734
Vitale G., et al., Biochem. Biophys. Res. Commun. 248, 706 (1998). (10.1006/bbrc.1998.9040) / Biochem. Biophys. Res. Commun. by Vitale G. (1998)
10.1042/bj3520739
J. O'Brien
Friedlander A., Dreier T., Ezzell J., Leppla S., Infect. Immun. 47, 306 (1985). (10.1128/iai.47.1.306-310.1985) / Infect. Immun. by Friedlander A. (1985)
Escuyer V., Collier R. J., Infect. Immun. 59, 3381 (1991). (10.1128/iai.59.10.3381-3386.1991) / Infect. Immun. by Escuyer V. (1991)
10.1073/pnas.89.21.10277
10.1038/385833a0
Milne J. C., Furlong D., Hanna P. C., Wall J. S., Collier R. J., J. Biol. Chem. 269, 20607 (1994). (10.1016/S0021-9258(17)32036-7) / J. Biol. Chem. by Milne J. C. (1994)
S. H. Leppla in Sourcebook of Bacterial Protein Toxins J. Alouf Ed. (Academic Press New York 1991) pp. 277–302.
Elliott J. L., Mogridge J., Collier R. J., Biochemistry 39, 6706 (2000). (10.1021/bi000310u) / Biochemistry by Elliott J. L. (2000)
Friedlander A. M., J. Biol. Chem. 261, 7123 (1986). (10.1016/S0021-9258(17)38364-3) / J. Biol. Chem. by Friedlander A. M. (1986)
Koehler T. M., Collier R. J., Mol. Microbiol. 5, 1501 (1991). (10.1111/j.1365-2958.1991.tb00796.x) / Mol. Microbiol. by Koehler T. M. (1991)
Walker B., Krishnasastry M., Zorn L., Bayley H., J. Biol. Chem. 267, 21782 (1992). (10.1016/S0021-9258(19)36680-3) / J. Biol. Chem. by Walker B. (1992)
Benson E. L., Huynh P. D., Finkelstein A., Collier R. J., Biochemistry 37, 3941 (1998). (10.1021/bi972657b) / Biochemistry by Benson E. L. (1998)
Miller C. J., Elliott J. L., Collier R. J., Biochemistry 38, 10432 (1999). (10.1021/bi990792d) / Biochemistry by Miller C. J. (1999)
Sellman B. R., Nassi S., Collier R. J., J. Biol. Chem. 276, 8371 (2001). (10.1074/jbc.M008309200) / J. Biol. Chem. by Sellman B. R. (2001)
Single-letter abbreviations for the amino acid residues are as follows: A Ala; C Cys; D Asp; E Glu; F Phe; G Gly; H His; I Ile; K Lys; L Leu; M Met; N Asn; P Pro; Q Gln; R Arg; S Ser; T Thr; V Val; W Trp; and Y Tyr.
Milne J. C., Blanke S. R., Hanna P. C., Collier R. J., Mol. Microbiol. 15, 661 (1995). (10.1111/j.1365-2958.1995.tb02375.x) / Mol. Microbiol. by Milne J. C. (1995)
Wesche J., Elliott J. L., Falnes P. O., Olsnes S., Collier R. J., Biochemistry 37, 15737 (1998). (10.1021/bi981436i) / Biochemistry by Wesche J. (1998)
Beauregard K. E., Collier R. J., Swanson J. A., Cell. Microbiol. 2, 251 (2000). (10.1046/j.1462-5822.2000.00052.x) / Cell. Microbiol. by Beauregard K. E. (2000)
B. R. Sellman M. Mourez R. J. Collier data not shown.
Ivins B. E., Ristroph J. D., Nelson G. O., Appl. Environ. Microbiol. 55, 2098 (1989). (10.1128/aem.55.8.2098-2100.1989) / Appl. Environ. Microbiol. by Ivins B. E. (1989)
Friedlander A. M., Pittman P. R., Parker G. W., JAMA 282, 2104 (1999). (10.1001/jama.282.22.2104) / JAMA by Friedlander A. M. (1999)
PA administered with Ribi Tri-Mix adjuvant elicits an anti-PA response and protects animals from challenge by B. anthracis spores (29). We tested the immunogenicity of the double deletion and F427A mutants by vaccinating groups of six male Fisher 344 rats intramuscularly at 0 3 and 6 weeks with each protein (50 μg) mixed with the Ribi Tri-Mix adjuvant (Sigma) in PBS. The animals were bled prior to the first injection and 14 days following each injection. On day 16 after the final injection the animals were challenged by intravenous injection of a lethal dose of WT-PA plus LF (30 μg + 6 μg respectively). Mock-vaccinated animals became moribund 74 min after injection whereas animals vaccinated with any of the DN mutants or with WT-PA survived without symptoms. Titers of PA-reactive and neutralizing antibodies elicited by the DN mutants (measured after the final injection) were greater than or equal to those elicited by WT-PA.
Y. Singh H. Kanna A. P. Chopra V. Mehra J. Biol. Chem. in press.
McBride B. W., et al., Vaccine 16, 810 (1998). (10.1016/S0264-410X(97)00268-5) / Vaccine by McBride B. W. (1998)
Supported by grant R37-AI22021 (R.J.C.) and training grant 5T32AI07410 (B.R.S.) from the NIH DSP/DGA (9860830006) (M.M.) and the Fondation Philippe (M.M.). R.J.C. has financial interest in AVANT Immunotherapeutics Inc.

Dates

Type	When
Created	23 years, 1 month ago (July 27, 2002, 5:52 a.m.)
Deposited	1 year, 7 months ago (Jan. 9, 2024, 11:26 p.m.)
Indexed	4 months, 3 weeks ago (April 10, 2025, 8:29 a.m.)
Issued	24 years, 4 months ago (April 27, 2001)
Published	24 years, 4 months ago (April 27, 2001)
Published Print	24 years, 4 months ago (April 27, 2001)

Funders 0

None

BibTeX

@article{Sellman_2001, title={Dominant-Negative Mutants of a Toxin Subunit: An Approach to Therapy of Anthrax}, volume={292}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.109563}, DOI={10.1126/science.109563}, number={5517}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Sellman, Bret R. and Mourez, Michael and John Collier, R.}, year={2001}, month=apr, pages={695–697} }

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