Properties of rat liver signal peptidase reconstituted into liposomes.
EDTA/KCl- or pyrophosphate-treated rough microsomes of rat liver clearly showed the co-translational cleavage of pre-human placental lactogen and translocation of the product into membrane vesicles. The signal peptidase fraction was isolated by chromatography on Sephacryl S-300 of deoxycholate-treated membranes and reconstituted into liposomes by dialysis or by the Biobeads SM-2 method. Assay of the signal peptidase activity was performed with pre-human placental lactogen synthesized by the reticulocyte lysate system programmed with human placental lactogen mRNA. The signal peptidase reconstituted into liposomes showed stable activity over the temperature range of 0 to 45 degrees C; in contrast, the detergent-solubilized signal peptidase of dog pancreatic membranes was completely inactivated at the unusually low temperature of 37 degrees C. It was shown that this inactivation was due to the presence of detergent. Signal peptidase from rat liver was insensitive to a variety of protease inhibitors, like the enzyme from dog pancreas, but differed from the latter in being inhibited by chymostatin and TPCK.
D000818:Animals; D002850:Chromatography, Gel; D003840:Deoxycholic Acid / Q000494:pharmacology; D004355:Drug Stability; D010450:Endopeptidases / Q000378:metabolism*; D005260:Female; D006358:Hot Temperature; D006801:Humans; D008081:Liposomes / Q000378:metabolism*; D008297:Male; D008565:Membrane Proteins; D008862:Microsomes, Liver / Q000201:enzymology*; D010920:Placenta / Q000032:analysis; D010928:Placental Lactogen / Q000235:genetics / Q000378:metabolism; D011247:Pregnancy; D011480:Protease Inhibitors / Q000494:pharmacology; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D012697:Serine Endopeptidases
D008081:Liposomes; D008565:Membrane Proteins; D011480:Protease Inhibitors; D011498:Protein Precursors; D012333:RNA, Messenger; D003840:Deoxycholic Acid; D010928:Placental Lactogen; D010450:Endopeptidases; D012697:Serine Endopeptidases; C026515:type I signal peptidase