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PMID: 14594844 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Structural and topographical studies of the type IV bundle-forming pilus assembly complex of enteropathogenic Escherichia coli.

Journal of bacteriology ·Vol. 185 ·No. 22 ·2003-11-00 ·Pages 6695-701

Hwang J, Bieber D, Ramer SW, Wu CY, Schoolnik GK

Abstract

The type IV bundle-forming pili (BFP) of enteropathogenic Escherichia coli (EPEC) are required for virulence in orally challenged human volunteers and for the localized adherence and autoaggregation in vitro phenotypes. BFP filament biogenesis and function are encoded by the 14-gene bfp operon. The BFP assembly complex, containing a BfpB-His6 fusion protein, was chemically cross-linked in situ, and the complex was then purified from BFP-expressing EPEC by a combination of nickel- and BfpB antibody-based affinity chromatography. Characterization of the isolated complex by immunoblotting using BFP protein-specific antibodies showed that at least 10 of the 14 proteins specified by the bfp operon physically interact to form an oligomeric complex. Proteins localized to the outer membrane, inner membrane, and periplasm are within this complex, thus demonstrating that the complex spans the periplasmic space. A combination of immunofluorescence and immuno-gold thin-section transmission electron microscopy studies localized this complex to one pole of the cell.

MeSH Terms
Cell Polarity Chromatography, Affinity Cross-Linking Reagents Escherichia coli/genetics,metabolism,ultrastructure Escherichia coli Proteins/chemistry,genetics,metabolism Fimbriae Proteins/chemistry,genetics,metabolism Fimbriae, Bacterial/metabolism Immunoblotting Microscopy, Electron Microscopy, Fluorescence Operon Recombinant Fusion Proteins/metabolism
Chemicals
Cross-Linking Reagents Escherichia coli Proteins Recombinant Fusion Proteins Fimbriae Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Hwang Jaiweon
Department of Medicine (Infectious Diseases and Geographic Medicine), Stanford Medical School, Stanford, California 94305, USA.
Bieber David
Ramer Sandra W
Wu Cheng-Yen
Schoolnik Gary K
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.185.22.6695-6701.2003
Published
2003-11-00
Pages
6695-701
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC262109
Subset
IM
Grants
NIAID NIH HHS · AI39521 · United States
External Links
PubMed source DOI: 10.1128/JB.185.22.6695-6701.2003 PMC Full Text View journal details
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