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Journal of Bacteriology, August 2001, p. 4848-4859, Vol. 183, No. 16
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.16.4848-4859.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Structure-Function Analysis of BfpB, a Secretin-Like Protein Encoded by the Bundle-Forming-Pilus Operon of Enteropathogenic Escherichia coli

Sarah A. Schmidt, David Bieber, Sandra W. Ramer,dagger Jaiweon Hwang, Cheng-Yen Wu, and Gary Schoolnik*

Departments of Medicine (Infectious Diseases and Geographic Medicine) and Microbiology & Immunology, Stanford Medical School, Stanford, California 94305

Received 2 February 2001/Accepted 22 May 2001

Production of type IV bundle-forming pili by enteropathogenic Escherichia coli (EPEC) requires BfpB, an outer-membrane lipoprotein and member of the secretin protein superfamily. BfpB was found to compose a ring-shaped, high-molecular-weight outer-membrane complex that is stable in 4% sodium dodecyl sulfate at temperatures of <= 65°C. Chemical cross-linking and immunoprecipitation experiments disclosed that the BfpB multimeric complex interacts with BfpG, and mutational studies showed that BfpG is required for the formation and/or stability of the multimer but not for the outer-membrane localization of BfpB. Formation of the BfpB multimer also does not require BfpA, the repeating subunit of the pilus filament. Functional studies of the BfpB-BfpG complex revealed that its presence confers vancomycin sensitivity, indicating that it may form an incompletely gated channel through the outer membrane. BfpB expression is also associated with accumulation of EPEC proteins in growth medium, suggesting that it may support both pilus biogenesis and protein secretion.

* Corresponding author. Mailing address: Beckman Center, Rm. 241, Stanford Medical School, Stanford, CA 94305. Phone: (650) 723-8158. Fax: (650) 723-1399. E-mail: schoolni{at}cmgm.stanford.edu.

dagger Present address: Maxygen, Redwood City, CA 94063.

Journal of Bacteriology, August 2001, p. 4848-4859, Vol. 183, No. 16
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.16.4848-4859.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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