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

► An ABC transporter and lipoprotein were functionally crosslinked in vitro. ► Two membrane subunits of LolCDE differ in function despite their similar structures. ► ATP hydrolysis is essential for transfer of a lipoprotein from LolE to LolA. A photo-sensitive amino acid analogue was introduced into...

Ausführliche Beschreibung

Gespeichert in:

Bibliographische Detailangaben
Veröffentlicht in:	FEBS letters 2013-01, Vol.587 (1), p.23-29
Hauptverfasser:	Mizutani, Masanori, Mukaiyama, Keita, Xiao, Jing, Mori, Makiko, Satou, Rika, Narita, Shin-ichiro, Okuda, Suguru, Tokuda, Hajime
Format:	Artikel
Sprache:	eng
Schlagworte:	ABC transporter Adenosine Triphosphate - metabolism ATP-Binding Cassette Transporters - chemistry ATP-Binding Cassette Transporters - genetics ATP-Binding Cassette Transporters - isolation & purification ATP-Binding Cassette Transporters - 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 Cytoplasmic membrane 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 Lipoprotein LolA Membrane subunit 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
Online-Zugang:	Volltext
Tags:	Tag hinzufügen Keine Tags, Fügen Sie den ersten Tag hinzu!

Beschreibung
Zusammenfassung:	► An ABC transporter and lipoprotein were functionally crosslinked in vitro. ► Two membrane subunits of LolCDE differ in function despite their similar structures. ► ATP hydrolysis is essential for transfer of a lipoprotein from LolE to LolA. 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. LolE and Palphysically interact by cross-linking study (View interaction) LolE and Palbind by cross-linking study (View interaction) Pal and Palbind by cross-linking study (View interaction) Pal and LolAphysically interact by competition binding (View interaction)
ISSN:	0014-5793 1873-3468
DOI:	10.1016/j.febslet.2012.11.009