Structural and functional characterization of OmpF porin mutants selected for larger pore size. I. Crystallographic analysis

J Biol Chem. 1996 Aug 23;271(34):20669-75.

Abstract

OmpF porin is a nonspecific pore protein from the outer membrane of Escherichia coli. Previously, a set of mutants was selected that allow the passage of long maltodextrins that do not translocate through the wild-type pore. Here, we describe the crystal structures of four point mutants and one deletion mutant from this set; their functional characterization is reported in the accompanying paper (Saint, N., Lou, K.-L., Widmer, C., Luckey, M., Schirmer, T., Rosenbusch, J. P. (1996) J. Biol. Chem. 271, 20676-20680). All mutations have a local effect on the structure of the pore constriction and result in a larger pore cross-section. Substitution of each of the three closely packed arginine residues at the pore constriction (Arg-42, Arg-82, and Arg-132) by shorter uncharged residues causes rearrangement of the adjacent basic residues. This demonstrates mutual stabilization of these residues in the wild-type porin. Deletion of six residues from the internal loop (Delta109-114) results in disorder of seven adjacent residues but does not alter the structure of the beta-barrel framework. Thus, the large hollow beta-barrel motif can be regarded as an autonomous structure.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacterial Outer Membrane Proteins / chemistry*
  • Bacterial Outer Membrane Proteins / genetics
  • Base Sequence
  • Crystallography, X-Ray
  • DNA Primers / chemistry
  • Escherichia coli
  • Hydrogen Bonding
  • Ion Channel Gating
  • Ion Channels / chemistry
  • Ion Channels / genetics
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Structure, Tertiary
  • Structure-Activity Relationship

Substances

  • Bacterial Outer Membrane Proteins
  • DNA Primers
  • Ion Channels

Associated data

  • PDB/1GFM
  • PDB/1GFN
  • PDB/1GFO
  • PDB/1GFP
  • PDB/1GFQ