Synthetic leader peptide modulates secretion of proteins from microinjected Xenopus oocytes.
To investigate the role of the leader peptide in modulating secretion from living cells, we injected a synthetic peptide into Xenopus oocytes. The peptide consisted of the NH2-terminal leader sequence of mouse immunoglobulin light chain precursor. We found that the leader peptide has two different roles in regulating secretion from the oocytes. First, it competitively inhibits the synthesis of secretory and membrane proteins but not of cytoplasmic proteins. The inhibition occurs both with oocyte proteins and with proteins directed by coinjected myeloma mRNA. The inhibition reaches a maximum 2 hr after injection and decays within 3 hr. It appears to be mediated through the cell membrane, because 125I-labeled leader peptide segregates into the membrane fraction of microinjected oocytes simultaneously with the interference with methionine incorporation. A second role of the microinjected leader peptide is to induce a rapid acceleration in the rate of export of secretory proteins from the oocyte. The maximal enhancement effect is obtained upon injection of 50 ng of leader peptide per oocyte. It is not merely due to the small size, negative charge, or hydrophobicity of the peptide, because enhanced secretion does not occur when glucagon, poly-L-glutamic acid, or Triton X-100 is injected. Furthermore, immunoreaction of the peptide with specific antibodies prior to microinjection prevents the accelerated export. Our observations indicate that in Xenopus oocytes, the leader peptide is involved in both translocation and later step(s) in the secretory pathway.
D000818:Animals; D004527:Egg Proteins / Q000235:genetics / Q000378:metabolism*; D005260:Female; D007700:Kinetics; D008715:Methionine / Q000378:metabolism; D008845:Microinjections; D009865:Oocytes / Q000187:drug effects / Q000502:physiology*; D010455:Peptides / Q000494:pharmacology*; D014176:Protein Biosynthesis / Q000187:drug effects; D021382:Protein Sorting Signals; D012333:RNA, Messenger / Q000235:genetics; D014981:Xenopus
D004527:Egg Proteins; D010455:Peptides; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D008715:Methionine