Protein disulfide-isomerase in rat exocrine pancreatic cells is exported from the endoplasmic reticulum despite possessing the retention signal.
Recently we found by immunogold electron microscopy that protein disulfide-isomerase (PDI), a major resident protein in the lumen of the endoplasmic reticulum (ER) of many cells, is exceptionally localized in rat exocrine pancreatic cells not only in the ER but also in plasma membranes and other organelles along secretory pathway (Akagi, S., Yamamoto, A., Yoshimori, T., Masaki, R., Ogawa, R., and Tashiro, Y. (1988) J. Histochem. Cytochem. 36, 1069-1074). These observations suggest that another type of PDI, e.g. one with a defective ER retention signal, might exist and be transported in the exocrine pancreatic cells. We therefore compared biochemical and immunochemical properties of the transported PDI with the authentic ER resident PDI. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, peptide mapping, urea-polyacrylamide gel electrophoresis, and isoelectric focusing showed that the former was indistinguishable from the latter. We prepared a polyclonal antibody against the synthetic hexapeptide, which corresponds to the carboxyl terminus of PDI containing the putative ER retention signal "KDEL." The epitopes of this antibody (anti-KDEL antibody) were located within the KDEL sequence. Anti-KDEL antibody reacted with PDI in both the plasma membranes and the ER of rat pancreatic cells in immunoblot analysis as well as in immunogold electron microscopy. These results suggest that PDI exported from the ER to the plasma membranes in rat exocrine pancreatic cells possesses the KDEL sequence.
Department of Physiology, Kansai Medical University, Osaka, Japan.
T
T
Yoshimori
T
T
Semba
H
H
Takemoto
S
S
Akagi
A
A
Yamamoto
Y
Y
Tashiro
D000591:Amino Acid Isomerases / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D000936:Antigen-Antibody Complex; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D004721:Endoplasmic Reticulum / Q000201:enzymology*; D005947:Glucose / Q000494:pharmacology; D006367:HeLa Cells / Q000187:drug effects / Q000201:enzymology; D006801:Humans; D015151:Immunoblotting; D019751:Intramolecular Transferases; D007700:Kinetics; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis; D010179:Pancreas / Q000201:enzymology* / Q000648:ultrastructure; D010449:Peptide Mapping; D021382:Protein Sorting Signals / Q000235:genetics*; D051381:Rats
D000936:Antigen-Antibody Complex; D009842:Oligopeptides; D021382:Protein Sorting Signals; D000591:Amino Acid Isomerases; D019751:Intramolecular Transferases; C021913:ornithine 5,4-aminomutase; D005947:Glucose