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 Pellett, S Articles by Welch, R A Search for Related Content PubMed PubMed Citation Articles by Pellett, S Articles by Welch, R A

 Previous Article  |  Next Article 

Infect Immun. 1990 March; 58(3): 822-827

Characterization of monoclonal antibodies against the Escherichia coli hemolysin.

Department of Medical Microbiology and Immunology, University of Wisconsin Medical School, Madison 53706.

ABSTRACT

Twelve monoclonal antibodies (MAbs) produced against the Escherichia coli hemolysin (HlyA) encoded by the hemolysin recombinant plasmid pWAM04 were studied. HlyA derivatives from recombinant strains with different plasmids encoding HlyA amino-terminal and carboxy-terminal truncates, HlyA in-frame deletions, and HlyA frameshift mutations were used in immunoblots to localize the antigenic determinants for the anti-HlyA MAbs. The mapping of the MAb epitopes was also facilitated by immunoblotting analysis of HlyA polypeptide fragments derived by cyanogen bromide cleavage. The HlyA epitopes for 11 of the MAbs were mapped to relatively small linear regions of the cytolysin ranging from 28 to 160 amino acids. Five of the MAbs (C10, G8, E2, B7, and D12) neutralized HlyA hemolytic activity to varying degrees. The epitopes for these neutralizing MAbs were found to reside within the following HlyA regions: C10 and G8, amino acids 2 to 160; E2, amino acids 161 to 194; B7, amino acids 518 to 598; and D12, amino acids 626 to 726. Hemolytically active HlyA was dependent on the action of the hlyC gene product. The D12 MAb recognized only HlyA produced by strains with an intact hlyC function. MAb A10 recognized an epitope within the HlyA region from amino acids 728 to 829 where a glycine-rich repeat domain exists; however, this MAb did not neutralize HlyA hemolytic activity. A HlyA domain map showing the anti-HlyA epitope location was constructed.

This article has been cited by other articles:

• Gleason, T. G., Houlgrave, C. W., May, A. K., Crabtree, T. D., Sawyer, R. G., Denham, W., Norman, J. G., Pruett, T. L. (1998). Hemolytically Active (Acylated) Alpha-Hemolysin Elicits Interleukin-1beta (IL-1beta ) but Augments the Lethality of Escherichia coli by an IL-1- and Tumor Necrosis Factor-Independent Mechanism. Infect. Immun. 66: 4215-4221 [Abstract] [Full Text]  
• Stanley, P., Koronakis, V., Hughes, C. (1998). Acylation of Escherichia coli Hemolysin: A Unique Protein Lipidation Mechanism Underlying Toxin Function. Microbiol. Mol. Biol. Rev. 62: 309-333 [Abstract] [Full Text]  
• Stanley, P, Packman, L., Koronakis, V, Hughes, C (1994). Fatty acylation of two internal lysine residues required for the toxic activity of Escherichia coli hemolysin. Science 266: 1992-1996 [Abstract]  
• Hackett, M, Guo, L, Shabanowitz, J, Hunt, D., Hewlett, E. (1994). Internal lysine palmitoylation in adenylate cyclase toxin from Bordetella pertussis. Science 266: 433-435 [Abstract]