PMID:8703042
| Citation |
Bochkareva, E, Seluanov, A, Bibi, E and Girshovich, A (1996) Chaperonin-promoted post-translational membrane insertion of a multispanning membrane protein lactose permease. J. Biol. Chem. 271:22256-61 |
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| Abstract |
Using an in vitro membrane-free translation system from Escherichia coli, it is shown that chaperonin GroEL added cotranslationally interacts with newly synthesized lactose permease (LacY), a polytopic membrane protein, thereby preventing aggregation. Subsequently, when the isolated GroEL-LacY complex is incubated with inverted membrane vesicles, the permease is inserted into the membrane in a MgATP-dependent manner. Post-translational membrane insertion is also observed when aggregation of newly synthesized LacY is prevented by addition of the nonionic detergent n-dodecyl-beta,D-maltoside during translation in place of GroEL. No membrane integration occurs with right-side-out vesicles, indicating that LacY interacts specifically only with the cytosolic face of the membrane. Ligand thiodigalactoside protection against alkylation of the Cys-148 residue in the permease shows proper post-translational insertion. Moreover, limited proteolysis of soluble LacY either complexed with GroEL or in detergent indicates that the newly synthesized protein assumes a conformation that is comparable to that of native, membrane-embedded permease prior to insertion into the membrane. |
| Links | |
| Keywords |
Adenosine Triphosphate/metabolism; Biological Transport; Chaperonin 60/metabolism; Cysteine/metabolism; Electrophoresis, Polyacrylamide Gel; Escherichia coli; Escherichia coli Proteins; Membrane Transport Proteins/metabolism; Monosaccharide Transport Proteins; Protein Processing, Post-Translational; Symporters |
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