A genetic approach for analysing surface-exposed regions of the OmpC protein of Escherichia coli K-12

Mol Microbiol. 1996 Feb;19(4):881-9. doi: 10.1046/j.1365-2958.1996.430957.x.

Abstract

Phage attachment sites on bacterial cell surfaces are provided by the exposed regions of outer membrane proteins and lipopolysaccharide (LPS). We have identified surface exposed residues of OmpC that are important for phage binding. This was accomplished by employing a genetic scheme in which two simultaneous selections enriched for ompC mutants defective in phage attachment, but retained functional channels. Mutational alterations were clustered in three regions of the OmpC protein. These regions also showed the greatest divergence from the analogous regions of the highly related OmpF and PhoE proteins. The majority of alterations (8 out of 11) occurred in a region of OmpC that is predicted to form a large exterior loop (loop 4). Interestingly, while the removal of this loop prevented phage binding, the deletion conferred enhanced channel activities. Another type of phage-resistant mutants synthesized defective LPS molecules. Biochemical analysis of mutant LPS revealed it to be of the Re-type LPS, lacking the heptose moieties from the LPS inner core. As a result of this LPS defect, many outer membrane proteins were present in somewhat reduced levels. The phage resistance seen in these mutants could be a result of both the presence of defective LPS and reduced OmpC levels.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Bacterial Outer Membrane Proteins / genetics
  • Bacterial Outer Membrane Proteins / metabolism*
  • Coliphages / metabolism*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Escherichia coli / virology
  • Ion Channels / metabolism
  • Molecular Sequence Data
  • Mutation
  • Sequence Analysis, DNA
  • Structure-Activity Relationship

Substances

  • Bacterial Outer Membrane Proteins
  • Ion Channels