Abstract
The relation of the structure of Pseudomonas aeruginosa exotoxin A (PA toxin) to its enzymatic activity (adenosine 5'-diphosphate-ribosyl transferase) in vitro and to its toxicity in vivo was examined. PA toxin is produced as a single polypeptide chain with a molecular weight of about 71,500. PA toxin is produced by Pseudomonas as a toxic proenzyme that lacks enzymatic activity. Adenosine 5'-diphosphate-ribosyl transferase activity is expressed when the molecule is denatured and reduced or when its is cleaved by Pseudomonas proteases to yield an enzymatically active 27,000-dalton fragment (fragment a). A 45,000-dalton protein is tentatively identified as the enzymatically inactive fragment b of PA toxin. Enzymatically active forms of the toxin lack toxicity for mouse L-cells or mouse lethality. Thus, it is concluded that the native toxin proenzyme is required for toxicity and that a structural rearrangement must precede its intracellular activity.
Full text
PDF![353](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9cd6/421528/f5aac4b953bd/iai00208-0365.png)
![354](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9cd6/421528/0250653fc284/iai00208-0366.png)
![355](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9cd6/421528/b94ea89b9021/iai00208-0367.png)
![356](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9cd6/421528/8250bb3e7048/iai00208-0368.png)
![357](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9cd6/421528/332efdbff6a8/iai00208-0369.png)
![358](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9cd6/421528/6c97be14f8c7/iai00208-0370.png)
![359](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9cd6/421528/a39ddd15e379/iai00208-0371.png)
![360](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9cd6/421528/c518b12f872c/iai00208-0372.png)
![361](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9cd6/421528/22bcb7e2f222/iai00208-0373.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- ALLEN E. H., SCHWEET R. S. Synthesis of hemoglobin in a cell-free system. I. Properties of the complete system. J Biol Chem. 1962 Mar;237:760–767. [PubMed] [Google Scholar]
- Atik M., Liu P. V., Hanson B. A., Amini S., Rosenberg C. F. Pseudomonas exotoxin shock. A preliminary report of studies in dogs. JAMA. 1968 Jul 15;205(3):134–140. doi: 10.1001/jama.205.3.134. [DOI] [PubMed] [Google Scholar]
- Callahan L. T., 3rd Purification and characterization of Pseudomonas aeruginosa exotoxin. Infect Immun. 1974 Jan;9(1):113–118. doi: 10.1128/iai.9.1.113-118.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Collier R. J. Diphtheria toxin: mode of action and structure. Bacteriol Rev. 1975 Mar;39(1):54–85. doi: 10.1128/br.39.1.54-85.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Collier R. J., Kandel J. Structure and activity of diphtheria toxin. I. Thiol-dependent dissociation of a fraction of toxin into enzymically active and inactive fragments. J Biol Chem. 1971 Mar 10;246(5):1496–1503. [PubMed] [Google Scholar]
- Finkelstein R. A., LoSpalluto J. J. Pathogenesis of experimental cholera. Preparation and isolation of choleragen and choleragenoid. J Exp Med. 1969 Jul 1;130(1):185–202. doi: 10.1084/jem.130.1.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gill D. M. Multiple roles of erythrocyte supernatant in the activation of adenylate cyclase by Vibrio cholerae toxin in vitro. J Infect Dis. 1976 Mar;133 (Suppl):55–63. doi: 10.1093/infdis/133.supplement_1.s55. [DOI] [PubMed] [Google Scholar]
- Iglewski B. H., Kabat D. NAD-dependent inhibition of protein synthesis by Pseudomonas aeruginosa toxin,. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2284–2288. doi: 10.1073/pnas.72.6.2284. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Iglewski B. H., Liu P. V., Kabat D. Mechanism of action of Pseudomonas aeruginosa exotoxin Aiadenosine diphosphate-ribosylation of mammalian elongation factor 2 in vitro and in vivo. Infect Immun. 1977 Jan;15(1):138–144. doi: 10.1128/iai.15.1.138-144.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kreger A. S., Griffin O. K. Physicochemical fractionation of extracellular cornea-damaging proteases of Pseudomonas aeruginosa. Infect Immun. 1974 May;9(5):828–834. doi: 10.1128/iai.9.5.828-834.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leppla S. H. Large-scale purification and characterization of the exotoxin of Pseudomonas aeruginosa. Infect Immun. 1976 Oct;14(4):1077–1086. doi: 10.1128/iai.14.4.1077-1086.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liu P. V. Exotoxins of Pseudomonas aeruginosa. I. Factors that influence the production of exotoxin A. J Infect Dis. 1973 Oct;128(4):506–513. doi: 10.1093/infdis/128.4.506. [DOI] [PubMed] [Google Scholar]
- Liu P. V. Exotoxins of Pseudomonas aeruginosa. I. Factors that influence the production of exotoxin A. J Infect Dis. 1973 Oct;128(4):506–513. doi: 10.1093/infdis/128.4.506. [DOI] [PubMed] [Google Scholar]
- Liu P. V. Extracellular toxins of Pseudomonas aeruginosa. J Infect Dis. 1974 Nov;130 (Suppl)(0):S94–S99. doi: 10.1093/infdis/130.supplement.s94. [DOI] [PubMed] [Google Scholar]
- Liu P. V. The roles of various fractions of Pseudomonas aeruginosa in its pathogenesis. 3. Identity of the lethal toxins produced in vitro and in vivo. J Infect Dis. 1966 Oct;116(4):481–489. doi: 10.1093/infdis/116.4.481. [DOI] [PubMed] [Google Scholar]
- Lui P. V. Biology of Pseudomonas aeruginosa. Hosp Pract. 1976 Jan;11(1):139–147. [PubMed] [Google Scholar]
- Pavlovskis O. R., Gordon F. B. Pseudomonas aeruginosa exotoxin: effect on cell cultures. J Infect Dis. 1972 Jun;125(6):631–636. doi: 10.1093/infdis/125.6.631. [DOI] [PubMed] [Google Scholar]
- Pavlovskis O. R., Voelker F. A., Shackelford A. H. Pseudomonas aeruginosa exotoxin in mice: histopathology and serum enzyme changes. J Infect Dis. 1976 Mar;133(3):253–259. doi: 10.1093/infdis/133.3.253. [DOI] [PubMed] [Google Scholar]
- Refsnes K., Olsnes S., Pihl A. On the toxic proteins abrin and ricin. Studies of their binding to and entry into Ehrlich ascites cells. J Biol Chem. 1974 Jun 10;249(11):3557–3562. [PubMed] [Google Scholar]
- Vasil M. L., Liu P. V., Iglewski B. H. Temperature-dependent inactivating factor of Pseudomonas aeruginosa exotoxin A. Infect Immun. 1976 May;13(5):1467–1472. doi: 10.1128/iai.13.5.1467-1472.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]