Translocation of proteins across the endoplasmic reticulum. I. Signal recognition protein (SRP) binds to in-vitro-assembled polysomes synthesizing secretory protein.
An 11S protein composed of six polypeptide chains was previously purified from a salt extract of dog pancreas microsomal membranes and shown to be required for translocation of nascent secretory protein across the microsomal membrane (Wistar and Blobel 1980 Proc. Natl. Acad. Sci. U. S. A. 77:7112-7116). This 11S protein, termed signal recognition protein (SRP), has been shown here (a) to inhibit translation in the wheat germ cell-free system selectively of mRNA for secretory protein (bovine preprolactin) but not of mRNA for cytoplasmic protein (alpha and beta chain of rabbit globin); (b) to bind with relatively low affinity (apparent KD less than 5 x 10(-5)) to monomeric wheat germ ribosomes; and (c) to bind selectively and with 6,000-fold higher affinity (apparent KD less than 8 x 10(-9)) to wheat germ ribosomes engaged in the synthesis of secretory protein but not to those engaged in the synthesis of cytoplasmic protein. Low- and high-affinity binding as well as the selective translation-inhibitory effect were abolished after modification of SRP by N-ethyl maleimide. High-affinity binding and the selective translation-inhibitory effect of SRP were largely abolished when the leucine (Leu) analogue beta-hydroxy leucine was incorporated into the nascent secretory polypeptide.
D000818:Animals; D004285:Dogs; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005914:Globins / Q000378:metabolism; D010179:Pancreas; D010455:Peptides / Q000378:metabolism* / Q000494:pharmacology; D011132:Polyribosomes / Q000378:metabolism*; D011388:Prolactin / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000187:drug effects; D011498:Protein Precursors / Q000378:metabolism*; D021382:Protein Sorting Signals; D012333:RNA, Messenger / Q000378:metabolism
D010455:Peptides; D011498:Protein Precursors; D021382:Protein Sorting Signals; D012333:RNA, Messenger; C017671:preprolactin; D011388:Prolactin; D005914:Globins