Diverse effects of phospholipids on lipoprotein sorting and ATP hydrolysis by the ABC transporter LolCDE complex.
The LolCDE complex of Escherichia coli releases outer membrane-specific lipoproteins from the inner membrane. Lipoproteins with Asp at +2 remain in the inner membrane since this residue functions as a LolCDE avoidance signal depending on phosphatidylethanolamine. We examined the effects of other phospholipids on lipoprotein sorting in proteoliposomes reconstituted with LolCDE and various synthetic phospholipids. The lipoprotein release and ATP hydrolysis were both low at 2 mM Mg(2+) but very high at 10 mM Mg(2+) in proteoliposomes containing cardiolipin alone. However, the Lol avoidance function was abolished at 10 mM Mg(2+), and the release of lipoproteins with Asp at +2 was as efficient as that of outer membrane-specific lipoproteins. The addition of phosphatidylethanolamine to cardiolipin stimulated the ATP hydrolysis and increased the Lol avoidance function of Asp at +2 at 2 mM Mg(2+). The addition of phosphatidylglycerol to cardiolipin nearly completely inhibited the release of lipoproteins with Asp at +2 even at 10 mM Mg(2+), while that of outer membrane-specific lipoproteins was not. Taken together, these results indicate that three major phospholipids of E. coli differently affect lipoprotein sorting and the activity of LolCDE.
Miyamoto
Shigehiko
S
Institute of Molecular and Cellular Biosciences, University of Tokyo, 1-1-1 Yayoi, Tokyo 113-0032, Japan.
Tokuda
Hajime
H
D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D002308:Cardiolipins / Q000378:metabolism / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism*; D008074:Lipoproteins / Q000187:drug effects / Q000378:metabolism*; D008081:Liposomes / Q000378:metabolism; D008274:Magnesium / Q000502:physiology; D018832:Molecular Chaperones; D033902:Periplasmic Binding Proteins / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000494:pharmacology; D010715:Phosphatidylglycerols / Q000494:pharmacology; D010743:Phospholipids / Q000494:pharmacology*; D021381:Protein Transport / Q000187:drug effects*; D011510:Proteolipids / Q000187:drug effects / Q000378:metabolism
D018528:ATP-Binding Cassette Transporters; D001425:Bacterial Outer Membrane Proteins; D002308:Cardiolipins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D008074:Lipoproteins; D008081:Liposomes; C116111:LolA protein, E coli; C087012:LolB protein, E coli; C407982:LolC protein, E coli; C407983:LolD protein, E coli; C407984:LolE protein, E coli; D018832:Molecular Chaperones; D033902:Periplasmic Binding Proteins; D010714:Phosphatidylethanolamines; D010715:Phosphatidylglycerols; D010743:Phospholipids; D011510:Proteolipids; C020485:proteoliposomes; C483858:phosphatidylethanolamine; D000255:Adenosine Triphosphate; D008274:Magnesium