The 70 carboxyl-terminal amino acids of nascent secretory proteins are protected from proteolysis by the ribosome and the protein translocation apparatus of the endoplasmic reticulum membrane.
We have used proteolysis to examine the environment through which nascent secretory proteins are translocated across the membrane of the endoplasmic reticulum. After solubilization of rough microsomes with detergent, fragments comprised of the approximately 70 carboxyl-terminal amino acids of translocating nascent chains initiated and targeted in vivo were protected from digestion by added proteases. About 40 amino acids of nascent chains were protected from proteolysis by the ribosome; thus, membrane-derived components protect an additional 30 amino acids. Under conditions in which those 30 additional amino acids are protected, only a small set of integral membrane proteins remained associated with the ribosome. These proteins include the Sec61 complex previously identified as the core component of the membrane-bound protein translocation apparatus. These results support the concept of a translocation pore that makes intimate contact with the ribosome and thereby protects nascent chains from proteolytic digestion for an additional, constant length.
Matlack
K E
KE
Department of Biochemistry and Biophysics, University of California Medical School, San Francisco 94143-0448.
Walter
P
P
D000818:Animals; D019826:Cholic Acid; D002793:Cholic Acids; D003902:Detergents; D004072:Digitonin; D004285:Dogs; D010450:Endopeptidases; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000378:metabolism; D008861:Microsomes / Q000378:metabolism*; D010179:Pancreas / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D011132:Polyribosomes / Q000378:metabolism; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism*; D012995:Solubility
D002793:Cholic Acids; D003902:Detergents; D008565:Membrane Proteins; D010446:Peptide Fragments; D011506:Proteins; D010450:Endopeptidases; D019826:Cholic Acid; D004072:Digitonin