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Journal of Bacteriology, December 2007, p. 9082-9089, Vol. 189, No. 24
0021-9193/07/$08.00+0     doi:10.1128/JB.01256-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Mapping Critical Interactive Sites within the Periplasmic Domain of the Vibrio cholerae Type II Secretion Protein EpsM{triangledown}

Tanya L. Johnson, Maria E. Scott,{dagger} and Maria Sandkvist*

Department of Microbiology and Immunology, University of Michigan Medical School, 1150 West Medical Center Drive, Ann Arbor, Michigan 48109

Received 3 August 2007/ Accepted 26 September 2007

The type II secretion (T2S) system is present in many gram-negative species, both pathogenic and nonpathogenic, where it supports the delivery of a variety of toxins, proteases, and lipases into the extracellular environment. In Vibrio cholerae, the T2S apparatus is composed of 12 Eps proteins that assemble into a multiprotein complex that spans the entire cell envelope. Two of these proteins, EpsM and EpsL, are key components of the secretion machinery present in the inner membrane. In addition to likely forming homodimers, EpsL and EpsM have been shown to form a stable complex in the inner membrane and to protect each other from proteolytic degradation. To identify and map the specific regions of EpsM involved in protein-protein interactions with both another molecule of EpsM and EpsL, we tested the interactions of deletion constructs of EpsM with full-length EpsM and EpsL by functional characterization and copurification as well as coimmunoprecipitation. Analysis of the truncated EpsM mutants revealed that the region of EpsM from amino acids 100 to 135 is necessary for EpsM to form homo-oligomers, while residues 84 to 99 appear to be critical for a stable interaction with EpsL.


* Corresponding author. Mailing address: Department of Microbiology and Immunology, University of Michigan Medical School, 1150 West Medical Center Drive, Ann Arbor, MI 48109. Phone: (734) 764-3552. Fax: (734) 764-3562. E-mail: mariasan{at}umich.edu

{triangledown} Published ahead of print on 5 October 2007.

{dagger} Present address: Western Michigan University, Kalamazoo, MI.


Journal of Bacteriology, December 2007, p. 9082-9089, Vol. 189, No. 24
0021-9193/07/$08.00+0     doi:10.1128/JB.01256-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.




This article has been cited by other articles:

  • Lybarger, S. R., Johnson, T. L., Gray, M. D., Sikora, A. E., Sandkvist, M. (2009). Docking and Assembly of the Type II Secretion Complex of Vibrio cholerae. J. Bacteriol. 191: 3149-3161 [Abstract] [Full Text]