This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Douglas, C M Articles by Collier, R J Search for Related Content PubMed PubMed Citation Articles by Douglas, C M Articles by Collier, R J

 Previous Article  |  Next Article 

J Bacteriol. 1987 November; 169(11): 4967-4971

Exotoxin A of Pseudomonas aeruginosa: substitution of glutamic acid 553 with aspartic acid drastically reduces toxicity and enzymatic activity.

Department of Microbiology, University of California, Los Angeles 90024.

ABSTRACT

Glutamic acid 553 of Pseudomonas aeruginosa exotoxin A (ETA) has been identified by photoaffinity labeling as a residue within the NAD binding site (S.F. Carroll and R.J. Collier, J. Biol. Chem. 262:8707-8711, 1987). To explore the function of Glu-553 we used oligonucleotide-directed mutagenesis to replace this residue with Asp in cloned ETA and expressed the mutant gene in Escherichia coli K-12. ADP-ribosylation activity of Asp-553 ETA in cell extracts was about 1,800-fold lower and toxicity for mouse L-M929 fibroblasts was at least 10,000-fold lower than that of the wild-type toxin. Extracts containing Asp-553 ETA inhibited the cytotoxicity of authentic ETA on L-M929 fibroblasts, suggesting that the mutant toxin competes for ETA receptors. The results indicate that Glu-553 is crucial for ADP-ribosylation activity and, consequently, cytotoxicity of ETA. Substitution or deletion of this residue may be a route to new ETA vaccines.

This article has been cited by other articles:

• Wykosky, J., Gibo, D. M., Debinski, W. (2007). A novel, potent, and specific ephrinA1-based cytotoxin against EphA2 receptor expressing tumor cells. Molecular Cancer Therapeutics 6: 3208-3218 [Abstract] [Full Text]  
• Onda, M., Nagata, S., FitzGerald, D. J., Beers, R., Fisher, R. J., Vincent, J. J., Lee, B., Nakamura, M., Hwang, J., Kreitman, R. J., Hassan, R., Pastan, I. (2006). Characterization of the B Cell Epitopes Associated with a Truncated Form of Pseudomonas Exotoxin (PE38) Used to Make Immunotoxins for the Treatment of Cancer Patients. J. Immunol. 177: 8822-8834 [Abstract] [Full Text]  
• Jorgensen, R., Yates, S. P., Teal, D. J., Nilsson, J., Prentice, G. A., Merrill, A. R., Andersen, G. R. (2004). Crystal Structure of ADP-ribosylated Ribosomal Translocase from Saccharomyces cerevisiae. J. Biol. Chem. 279: 45919-45925 [Abstract] [Full Text]  
• Watanabe, M., Kono, T., Matsushima-Hibiya, Y., Kanazawa, T., Nishisaka, N., Kishimoto, T., Koyama, K., Sugimura, T., Wakabayashi, K. (1999). Molecular cloning of an apoptosis-inducing protein, pierisin, from cabbage butterfly: Possible involvement of ADP-ribosylation in its activity. Proc. Natl. Acad. Sci. USA 96: 10608-10613 [Abstract] [Full Text]  
• Marsischky, G. T., Wilson, B. A., Collier, R. J. (1995). Role of Glutamic Acid 988 of Human Poly-ADP-ribose Polymerase in Polymer Formation. J. Biol. Chem. 270: 3247-3254 [Abstract] [Full Text]  
• Han, X. Y., Galloway, D. R. (1995). Active Site Mutations of Pseudomonas aeruginosa Exotoxin A. J. Biol. Chem. 270: 679-684 [Abstract] [Full Text]  
• Burnette, W., Cieplak, W, Mar, V., Kaljot, K., Sato, H, Keith, J. (1988). Pertussis toxin S1 mutant with reduced enzyme activity and a conserved protective epitope. Science 242: 72-74 [Abstract]