title: Functional differentiation of structurally similar membrane subunits of the ABC transporter LolCDE complex.
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.
authors.affiliation: Faculty of Nutritional Sciences, University of Morioka, Takizawa, Japan.
authors.forename: Masanori
authors.initials: M
authors.lastname: Mizutani
authors.forename: Keita
authors.initials: K
authors.lastname: Mukaiyama
authors.forename: Jing
authors.initials: J
authors.lastname: Xiao
authors.forename: Makiko
authors.initials: M
authors.lastname: Mori
authors.forename: Rika
authors.initials: R
authors.lastname: Satou
authors.forename: Shin-ichiro
authors.initials: S
authors.lastname: Narita
authors.forename: Suguru
authors.initials: S
authors.lastname: Okuda
authors.forename: Hajime
authors.initials: H
authors.lastname: Tokuda
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001577:Benzophenones / Q000737:chemistry; D015153:Blotting, Western; D002846:Chromatography, Affinity; D018388:Codon, Terminator; D003432:Cross-Linking Reagents / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D057927:Hydrophobic and Hydrophilic Interactions; D008024:Ligands; D046912:Multiprotein Complexes / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000737:chemistry; D055668:Photochemical Processes; D054730:Protein Interaction Domains and Motifs; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: C488060:4-benzoylphenylalanine; D018528:ATP-Binding Cassette Transporters; D001425:Bacterial Outer Membrane Proteins; D001577:Benzophenones; D018388:Codon, Terminator; D003432:Cross-Linking Reagents; D029968:Escherichia coli Proteins; D008024:Ligands; C407982:LolC protein, E coli; C407983:LolD protein, E coli; C407984:LolE protein, E coli; D046912:Multiprotein Complexes; D050505:Mutant Proteins; D021122:Protein Subunits; D011994:Recombinant Proteins; D010649:Phenylalanine; D000255:Adenosine Triphosphate