Protein insertion into the endoplasmic reticulum of permeabilized cells.
We have established efficient translocation of newly synthesized proteins into the endoplasmic reticulum of permeabilized Mel Juso cells. By site-specific photo-crosslinking we show that translocating polypeptide chains contact the same components of permeabilized cells ER as in dog pancreas rough microsomes. This cellular assay system has the potential to overcome the limitations of isolated microsomes in investigating the molecular environment of a newly synthesized protein after they have left the ER translocation site.
Jadot
M
M
Zentrum für Molekulare Biologie, Universität Heidelberg (ZMBH), Germany.
Hofmann
M W
MW
Graf
R
R
Quader
H
H
Martoglio
B
B
D000818:Animals; D001692:Biological Transport; D002460:Cell Line; D002463:Cell Membrane Permeability; D003062:Codon; D003432:Cross-Linking Reagents; D004285:Dogs; D019286:Endopeptidase K; D004721:Endoplasmic Reticulum / Q000378:metabolism* / Q000648:ultrastructure; D006031:Glycosylation; D007425:Intracellular Membranes / Q000378:metabolism; D008027:Light; D008565:Membrane Proteins / Q000378:metabolism*; D017830:Octoxynol; D010179:Pancreas / Q000648:ultrastructure; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D012503:Saponins / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000494:pharmacology; D012995:Solubility
D003062:Codon; D003432:Cross-Linking Reagents; D008565:Membrane Proteins; D012333:RNA, Messenger; D012503:Saponins; D017830:Octoxynol; D012697:Serine Endopeptidases; D019286:Endopeptidase K