Solubilization and functional reconstitution of the protein-translocation enzymes of Escherichia coli.
The SecY protein and other membrane proteins of Escherichia coli were solubilized by mixed micelles of n-octyl beta-D-glucopyranoside, phospholipids, and glycerol. Proteoliposomes formed from this extract by detergent dialysis supported energy-dependent translocation and processing of pro-OmpA. Translocation required ATP, SecY, and SecA and was stimulated by a proton-motive force. These results provide an important assay for the isolation and identification of membrane components involved in protein translocation.
Driessen
A J
AJ
Molecular Biology Institute, University of California, Los Angeles 90024-1737.
Wickner
W
W
D001425:Bacterial Outer Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism; D001426:Bacterial Proteins / Q000302:isolation & purification* / Q000378:metabolism; D001436:Bacteriorhodopsins / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology*; D002850:Chromatography, Gel; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins; D007700:Kinetics; D008081:Liposomes; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D011510:Proteolipids / Q000378:metabolism; D006180:Proton-Translocating ATPases / Q000302:isolation & purification* / Q000378:metabolism; D000069816:SEC Translocation Channels; D012995:Solubility
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D008081:Liposomes; D008565:Membrane Proteins; D011510:Proteolipids; D000069816:SEC Translocation Channels; C057841:SecY protein, E coli; C020485:proteoliposomes; D001436:Bacteriorhodopsins; D006180:Proton-Translocating ATPases