Functional differentiation of structurally similar membrane subunits of the ABC transporter LolCDE complex

FEBS Lett. 2013 Jan 4;587(1):23-9. doi: 10.1016/j.febslet.2012.11.009. Epub 2012 Nov 24.

Abstract

A photo-sensitive amino acid analogue was introduced into an outer membrane lipoprotein, Pal, and then subjected to photo-crosslinking with the lipoprotein-specific ABC transporter LolCDE. Pal crosslinked to LolE but not LolC in vivo despite that both are structurally similar membrane subunits. LolCDE liganded with Pal containing the photo-sensitive amino acid analogue was isolated and subjected to in vitro photo-crosslinking. LolE was found to be the binding site for Pal. ATP binding to LolD decreased the LolE-Pal crosslinking by decreasing their hydrophobic interaction. ATP hydrolysis in the presence of LolA completely abolished the LolE-Pal crosslinking and, concomitantly, generated a new LolA-Pal crosslinked product.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP-Binding Cassette Transporters / chemistry
  • ATP-Binding Cassette Transporters / genetics
  • ATP-Binding Cassette Transporters / isolation & purification
  • ATP-Binding Cassette Transporters / metabolism*
  • Adenosine Triphosphate / metabolism
  • Bacterial Outer Membrane Proteins / chemistry
  • Bacterial Outer Membrane Proteins / genetics
  • Bacterial Outer Membrane Proteins / isolation & purification
  • Bacterial Outer Membrane Proteins / metabolism*
  • Benzophenones / chemistry
  • Blotting, Western
  • Chromatography, Affinity
  • Codon, Terminator
  • Cross-Linking Reagents / chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / isolation & purification
  • Escherichia coli Proteins / metabolism*
  • Hydrophobic and Hydrophilic Interactions
  • Ligands
  • Multiprotein Complexes / chemistry
  • Multiprotein Complexes / genetics
  • Multiprotein Complexes / isolation & purification
  • Multiprotein Complexes / metabolism*
  • Mutagenesis, Site-Directed
  • Mutant Proteins / chemistry
  • Mutant Proteins / metabolism
  • Phenylalanine / analogs & derivatives
  • Phenylalanine / chemistry
  • Photochemical Processes
  • Protein Interaction Domains and Motifs
  • Protein Subunits / chemistry
  • Protein Subunits / genetics
  • Protein Subunits / isolation & purification
  • Protein Subunits / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism

Substances

  • 4-benzoylphenylalanine
  • ATP-Binding Cassette Transporters
  • Bacterial Outer Membrane Proteins
  • Benzophenones
  • Codon, Terminator
  • Cross-Linking Reagents
  • Escherichia coli Proteins
  • Ligands
  • LolC protein, E coli
  • LolD protein, E coli
  • LolE protein, E coli
  • Multiprotein Complexes
  • Mutant Proteins
  • Protein Subunits
  • Recombinant Proteins
  • Phenylalanine
  • Adenosine Triphosphate