Reconstitution of protein translocation activity from partially solubilized microsomal vesicles.
We have used a reconstitution assay to demonstrate that protein translocation activity can be recovered after microsomal vesicles derived from the rough endoplasmic reticulum have been partially solubilized with n-octyl-beta-glucopyranoside. Two independent approaches were used to establish conditions for partially solubilizing microsomal membranes. When the lipid bilayer was disrupted by detergents to the extent that the integrity of the lipid bilayer had been perturbed, membranes were inactive for translocation. However, detergent-treated membranes could be reconstituted in good yield into a translocation competent form once the detergent was removed.
Zimmerman
D L
DL
Department of Biochemistry and Biophysics, University of California, San Francisco 94143-0448.
Walter
P
P
D000818:Animals; D002458:Cell Fractionation; D003902:Detergents / Q000494:pharmacology; D004285:Dogs; D004492:Edetic Acid / Q000494:pharmacology; D010450:Endopeptidases / Q000302:isolation & purification / Q000378:metabolism; D005960:Glucosides / Q000494:pharmacology; D007425:Intracellular Membranes / Q000378:metabolism*; D007700:Kinetics; D008565:Membrane Proteins; D008861:Microsomes / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000235:genetics*; D012697:Serine Endopeptidases; D012995:Solubility
D003902:Detergents; D005960:Glucosides; D008565:Membrane Proteins; D011506:Proteins; C018619:octyl-beta-D-glucoside; D004492:Edetic Acid; D010450:Endopeptidases; D012697:Serine Endopeptidases; C026515:type I signal peptidase