Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Science 25 February 1972:
Vol. 175. no. 4024, pp. 901 - 903
DOI: 10.1126/science.175.4024.901
Prev | Table of Contents | Next

Articles

Reconstitution of Diphtheria Toxin from Two Nontoxic Cross-Reacting Mutant Proteins

Tsuyoshi Uchida 1, A. M. Pappenheimer Jr. 1, and Annabel Avery Harper 1

1 Biological Laboratories, Harvard University, Cambridge, Massachusetts 02138

The isolation of a new type of mutant Corynephage beta, which carries a missense mutation in the structural gene for diphtheria toxin synthesis is described. The lysogenic C78(beta197)tox-crm+ strain of Corynebacterium diphtheriae produces a nontoxic, extracellular protein of molecular weight 62,000. This protein is immunologically indistinguishable from toxin itself but inhibits the action of toxin on HeLa cells, probably by competing for attachment sites on the cell membrane. In contrast to fragment A derived from diphtheria toxin, fragment A197 is unable to catalyze the inactivation of eucaryotic polypeptidyl-transfer RNA-transferase II. When mixtures of the two nontoxic mutant proteins, enzymically active crm45 protein and enzymically inactive crm197 protein, are subjected to mild treatment with trypsin in the presence of a thiol and then allowed to reoxidize after dialysis to remove excess thiol, "diphtheria toxin" is reconstituted in high yield.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Endosome fusion induced by diphtheria toxin translocation domain.
A. Antignani and R. J. Youle (2008)
PNAS 105, 8020-8025
   Abstract »    Full Text »    PDF »
Diphtheria Toxin Mutant CRM197 Possesses Weak EF2-ADP-ribosyl Activity that Potentiates its Anti-tumorigenic Activity.
T. Kageyama, M. Ohishi, S. Miyamoto, H. Mizushima, R. Iwamoto, and E. Mekada (2007)
J. Biochem. 142, 95-104
   Abstract »    Full Text »    PDF »
Antigen Processing of the Heptavalent Pneumococcal Conjugate Vaccine Carrier Protein CRM197 Differs Depending on the Serotype of the Attached Polysaccharide.
E. G. Leonard, D. H. Canaday, C. V. Harding, and J. R. Schreiber (2003)
Infect. Immun. 71, 4186-4189
   Abstract »    Full Text »    PDF »
CpG Oligodeoxynucleotides Act as Adjuvants for Pneumococcal Polysaccharide-Protein Conjugate Vaccines and Enhance Antipolysaccharide Immunoglobulin G2a (IgG2a) and IgG3 Antibodies.
R. S. Chu, T. McCool, N. S. Greenspan, J. R. Schreiber, and C. V. Harding (2000)
Infect. Immun. 68, 1450-1456
   Abstract »    Full Text »    PDF »
B- and T-Cell Immune Responses to Pneumococcal Conjugate Vaccines: Divergence between Carrier- and Polysaccharide-Specific Immunogenicity.
T. L. McCool, C. V. Harding, N. S. Greenspan, and J. R. Schreiber (1999)
Infect. Immun. 67, 4862-4869
   Abstract »    Full Text »    PDF »
{gamma}3 Gene-Disrupted Mice Selectively Deficient in the Dominant IgG Subclass Made to Bacterial Polysaccharides Undergo Normal Isotype Switching After Immunization with Polysaccharide-Protein Conjugate Vaccines.
D. A. Shapiro, D. S. Threadgill, M. J. Copfer, D. A. Corey, T. L. McCool, L. L. McCormick, T. R. Magnuson, N. S. Greenspan, and J. R. Schreiber (1998)
J. Immunol. 161, 3393-3399
   Abstract »    Full Text »    PDF »
Diphtheria Toxin Translocation across Endosome Membranes. A NOVEL CELL PERMEABILIZATION ASSAY REVEALS NEW DIPHTHERIA TOXIN FRAGMENTS IN ENDOCYTIC VESICLES.
T. Umata and E. Mekada (1998)
J. Biol. Chem. 273, 8351-8359
   Abstract »    Full Text »    PDF »
Cell-mediated Reduction and Incomplete Membrane Translocation of Diphtheria Toxin Mutants with Internal Disulfides in the A Fragment.
P.Ø. Falnes and S. Olsnes (1995)
J. Biol. Chem. 270, 20787-20793
   Abstract »    Full Text »    PDF »
Effect of Carrier Protein Priming on Antibody Responses to Haemophilus influenzae Type b Conjugate Vaccines in Infants.
D. M. Granoff, S. J. Holmes, R. B. Belshe, M. T. Osterholm, J. E. McHugh, and E. L. Anderson (1994)
JAMA 272, 1116-1121
   Abstract »    PDF »
Mutations in diphtheria toxin separate binding from entry and amplify immunotoxin selectivity.
L Greenfield, V. Johnson, and R. Youle (1987)
Science 238, 536-539
   Abstract »    PDF »
Peripheral Nerve Segmental Demyelination Induced by Intraneural Diphtheria Toxin Injection: Sodium Na 24 and Carbon 14-Labeled Inulin Kinetics in Diphtheria Toxin-Injected Nerve and the Effect of Hydrocortisone.
V. A. Levin, E. Chelmicka-Szorc, and B. G. W. Arnason (1974)
Arch Neurol 30, 163-168
   Abstract »    PDF »
Diphtheria: Recent studies have clarified the molecular mechanisms involved in its pathogenesis.
A. M. Pappenheimer Jr. and D. M. Gill (1973)
Science 182, 353-358
   PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)