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Title: Associations of Escherichia coli K-12 OmpF trimers with rough and smooth lipopolysaccharides

Abstract

The associations of both rough and smooth lipopolysaccharides (LPS) with the OmpF porin of Escherichia coli K-12 were examined in galE strains deleted for ompC. Transformation with pSS37 and growth with galactose conferred the ability to assemble a Shigella dysenteriae O antigen onto the core oligosaccharide of E. coli K-12 LPS. The association of LPS with OmpF trimers was assessed by staining, autoradiography of LPS specifically labeled with (1-14C)galactose, and Western immunoblotting with a monoclonal antibody specific for OmpF trimers. These techniques revealed that the migration distances and multiple banding patterns of OmpF porin trimers in sodium dodecyl sulfate-polyacrylamide gels were dictated by the chemotype of associated LPS. Expression of smooth LPS caused almost all of the trimeric OmpF to run in gels with a slower mobility than trimers from rough strains. The LPS associated with trimers from a smooth strain differed from the bulk-phase LPS by consisting almost exclusively of molecules with O antigen.

Authors:
; ;  [1]
  1. Louisiana State Univ. Medical Center, New Orleans (USA)
Publication Date:
OSTI Identifier:
6511251
Resource Type:
Journal Article
Journal Name:
Journal of Bacteriology; (USA)
Additional Journal Information:
Journal Volume: 172:9; Journal ID: ISSN 0021-9193
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ESCHERICHIA COLI; GROWTH; LIPOPOLYSACCHARIDES; AUTORADIOGRAPHY; CARBON 14 COMPOUNDS; PORINS; TWO-DIMENSIONAL ELECTROPHORESIS; BACTERIA; CARBOHYDRATES; ELECTROPHORESIS; LABELLED COMPOUNDS; LIPIDS; MEMBRANE PROTEINS; MICROORGANISMS; ORGANIC COMPOUNDS; POLYSACCHARIDES; PROTEINS; SACCHARIDES; 550201* - Biochemistry- Tracer Techniques; 550701 - Microbiology- Tracer Techniques

Citation Formats

Diedrich, D L, Stein, M A, and Schnaitman, C A. Associations of Escherichia coli K-12 OmpF trimers with rough and smooth lipopolysaccharides. United States: N. p., 1990. Web.
Diedrich, D L, Stein, M A, & Schnaitman, C A. Associations of Escherichia coli K-12 OmpF trimers with rough and smooth lipopolysaccharides. United States.
Diedrich, D L, Stein, M A, and Schnaitman, C A. Sat . "Associations of Escherichia coli K-12 OmpF trimers with rough and smooth lipopolysaccharides". United States.
@article{osti_6511251,
title = {Associations of Escherichia coli K-12 OmpF trimers with rough and smooth lipopolysaccharides},
author = {Diedrich, D L and Stein, M A and Schnaitman, C A},
abstractNote = {The associations of both rough and smooth lipopolysaccharides (LPS) with the OmpF porin of Escherichia coli K-12 were examined in galE strains deleted for ompC. Transformation with pSS37 and growth with galactose conferred the ability to assemble a Shigella dysenteriae O antigen onto the core oligosaccharide of E. coli K-12 LPS. The association of LPS with OmpF trimers was assessed by staining, autoradiography of LPS specifically labeled with (1-14C)galactose, and Western immunoblotting with a monoclonal antibody specific for OmpF trimers. These techniques revealed that the migration distances and multiple banding patterns of OmpF porin trimers in sodium dodecyl sulfate-polyacrylamide gels were dictated by the chemotype of associated LPS. Expression of smooth LPS caused almost all of the trimeric OmpF to run in gels with a slower mobility than trimers from rough strains. The LPS associated with trimers from a smooth strain differed from the bulk-phase LPS by consisting almost exclusively of molecules with O antigen.},
doi = {},
journal = {Journal of Bacteriology; (USA)},
issn = {0021-9193},
number = ,
volume = 172:9,
place = {United States},
year = {1990},
month = {9}
}