Summary
The transcellular movement of fucosylated glycoproteins has been studied in vitro using rat pancreatic lobules and cell fractionation procedures, and has been compared with the well established pathway of secretory proteins. Using tritiated leucine as pulse label for the latter, their translocation from the rough endoplasmatic reticulum into the Golgi complex and finally into zymogen granules could be followed. In the case of glycoproteins, 14C-fucose was incorporated mainly into the smooth microsomal fraction (representative of the Golgi complex) and only one third of this specific activity was transported into the zymogen granule fraction. A detailed analysis of this fraction after separation of the content of zymogen granules from their membranes revealed a predominant labeling of membrane glycoproteins by 14C-fucose. In comparison, leucine-labeled bulk proteins were found almost exclusively in the zymogen granule content fraction, with little radioactivity in the membrane fraction. The data indicate a concomitant synthesis of fucosylated glycoproteins destined in part for the zymogen granule membrane and to a greater amount associated with the smooth microsomal fraction. The results are discussed in the light of recent findings indicating that about 40% of the proteins in the zymogen granule membrane are made up of one major glycoprotein which could be involved in the mechanism of exocytosis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bennett, G., Leblond, C.P., Haddad, A.: Migration of glycoprotein from the Golgi apparatus to the cell surface of various cell types as shown by radioautography after labeled fucose injection into rats. J. Cell Biol. 60, 258–284 (1974)
Bernfeld, P.: Amylases, α and β. In: Methods in enzymology (S.P. Colowick and N.O. Kaplan, eds.), Vol. 1, pp. 149–158. New York: Academic Press 1955
Bieger, W., Seybold, J., Kern, H.F.: Studies on intracellular transport of secretory proteins in the rat exocrine pancreas. V. Kinetic studies on accelerated transport following caerulein infusion in vivo. Cell Tiss. Res. 170, 203–219 (1976)
Castle, J.D., Palade, G.E.: Secretion granules of the rabbit parotid. Selective removal of secretory contaminants from granule membranes. J. Cell Biol. 76, 323–340 (1978)
Chen, P.S., Toribara, T.Y., Warner, H.: Microdetermination of phosphorus. Analyt. Chem. 28, 1756–1758 (1956)
Cook, G.M.W.: Biosynthesis of plasma membrane glycoproteins. In: cell surface reviews (G. Poste and G.L. Nicolson, eds.), Vol. IV. Amsterdam: North Holland Publ. Co. 1978
Cooperstein, S.J., Lazarow, A.: A microspectrophotometric method for the determination of cytochrome oxidase. J. biol. Chem. 189, 665–670 (1951)
Fambrough, D.M., Devreotes, P.N.: Newly synthesized acetylcholine receptors are located in the Golgi apparatus. J. Cell Biol. 76, 237–244 (1978)
Folk, J.E., Piez, K.A., Carroll, W.R., Gladner, J.A.: Carboxypeptidase B, IV. J. biol. Chem. 235, 2272–2277 (1960)
Hatcher, D.W., Goldstein, G.: Improved methods for determination of RNA and DNA. Analyt. Biochem. 31, 42–50 (1969)
Hummel, B.C.: A modified spectrophotometric determination of chymotrypsin, trypsin and thrombin. Canad. J. Biochem. 37, 1393–1399 (1959)
Jamieson, J.D.: Transport and discharge of exportable proteins in pancreatic exocrine cells: in vitro studies. In: Current topics in membranes and transport (F. Bronner and A. Kleinzeller, eds.), Vol. 3, pp. 273–338. New York: Academic Press Inc. 1972
Kronquist, K.E., Elmahdy, A., Ronzio, R.A.: Synthesis and subcellular distribution of heparan sulfate in the rat exocrine pancreas. Arch. Biochem. Biophys. 182, 188–196 (1977)
Leblond, C.P., Bennett, G.: Role of the Golgi apparatus in terminal glycosylation. In: International cell biology (B.R., Brinkley and K.R. Porter, eds.). New York: Rockefeller Univ. Press 1977
Lewis, D.S., MacDonald, R.J., Kronquist, K.E., Ronzio, R.A.: Purification and partial characterization of an integral membrane glycoprotein from zymogen granules of dog pancreas. FEBS Letters 76, 115–120 (1977)
Lowry, O.H., Rosenbrough, N.J., Farr, A.L., Randell, R.J.: Protein measurement with the Folin phenol reagent. J. biol. Chem. 193, 265–275 (1951)
Meldolesi, J., Borgese, N., DeCamilli, P., Ceccarelli, B.: Cytoplasmic membranes in the secretory process. In: Cell surface reviews (G. Poste and G.L. Nicolson, eds.), Vol. V. Amsterdam: North Holland Publ. Co. 1978
Michaels, J.E., Leblond, C.P.: Transport of glycoprotein from Golgi apparatus to cell surface by means of carrier vesicles, as shown by radioautography of mouse colonic epithelium after injection of 3H-fucose. J. Microsc. Biol. Cell. 25, 243–248 (1976)
Morré, D.J., Ovtracht, L.: Dynamics of the Golgi apparatus: membrane differentiation and membrane flow. In: Int. Review Cytology (G.H. Bourne, J.F., Danielli and K.W. Jeon, eds.), Suppl. 5. New York: Academic Press 1977
Reggio, H., Palade, G.E.: Sulfated compounds in the secretion and zymogen granule content of the guinea pig pancreas. J. Cell Biol. 70, 360a (1976)
Ronzio, R.A.: glycoprotein synthesis in the adult rat pancreas. I. Subcellular distribution of uridine diphosphate galactose: glycoprotein galactosyltransferase and thiamine pyrophosphate phosphohydrolase. Biochim. biophys. Acta (Amst.) 313, 286–295 (1973a)
Ronzio, R.A.: Glycoprotein synthesis in the adult rat pancreas. II. Characterization of Golgi rich fractions. Arch. Biochem. Biophys. 159, 777–784 (1973b)
Ronzio, R.A., Mohrlock, S.H.: A possible role of Golgi membrane-associated galactosyltransferase in the formation of zymogen granule glycoproteins Arch. Biochem. Biophys. 181, 128–136 (1977)
Ronzio, R.A., Kronquist, K.E., Lewis, D.S., MacDonald, R.J., Mohrlock, S.H., O'Donnell, J.J.: Glycoprotein synthesis in the adult rat pancreas. IV. Subcellular distribution of membrane glycoproteins. Biochim. biophys. Acta (Amst.) 508, 65–84 (1978)
Sandoz, D., Roland, J.C.: Radioautographic study of glycoproteins and polyglycans. J. Microscop. Biol. Cell 27, 207–214 (1976)
Schachter, H.: The subcellular sites of glycosylation. Biochem. Soc. Symp. 40, 50–71 (1974)
Scheele, G.A.: Two-dimensional gel analysis of soluble proteins. Characterization of guinea pig exocrine pancreatic proteins. J. biol. Chem. 250, 5375–5385 (1975)
Scheele, G.A., Palade, G.E.: Studies on the guinea pig pancreas. Parallel discharge of exocrine enzyme activities. J. biol. Chem. 250, 2660–2670 (1975)
Scheele, G.A., Palade, G.E., Tartakoff, A.M.: Cell fractionation on the guinea pig pancreas. Redistribution of exocrine proteins during tissue homogenisation. J. cell Biol. (in press)
Tartakoff, A.M., Greene, L.J., Palade, G.E.: Studies on the guinea pig pancreas. Fractionation and partial characterization of exocrine proteins. J. biol. Chem. 249, 7420–7432 (1974)
Tartakoff, A.M., Jamieson, J.D.: Subcellular fractionation of the pancreas. In: Methods in enzymology (S.S. Fleischer and L. Packer, eds.), Vol. 31, pp. 41–59. New York: Academic Press 1974
Völkl, A., Bieger, W., Kern, H.F.: Studies on secretory glycoproteins in the rat exocrine pancreas. I. Fine structure of the Golgi complex and release of fucose-labeled proteins after in vivo stimulation with Caerulein. Cell Tiss. Res. 175, 227–243 (1976)
Völkl, A., Seybold, J., Kern, H.F.: Studies on secretory glycoproteins in the rat exocrine pancreas. II. Identification of a fetuin: fucosyl transferase in the smooth microsomal fraction. Cell Tiss. Res. 186, 111–119 (1978)
Whaley, W.G.: The Golgi apparatus. Cell biology monographs, Vol. 2. Berlin-Heidelberg-New York: Springer 1975
Zinterhofer, L., Wardlaw, S., Jatlow, P., Seligson, D.: Nephelometric determination of pancreatic enzymes. II. Lipase. Clin. chim. Acta 44, 173–178 (1973)
Author information
Authors and Affiliations
Additional information
Dedicated to Professor Gerhard Petry, Marburg, on the occasion of his 65th birthday
Supported by a grant from the Deutsche Forschungsgemeinschaft, Bonn-Bad Godesberg (SFB 122, project C5). The expert technical assistance of Mrs. H. Beil and Mrs. U. Kubik and the editorial help of Mrs. G. Lesch is gratefully acknowledged
Rights and permissions
About this article
Cite this article
Völkl, A., Schick, J., Adler, G. et al. Studies on secretory glycoproteins in the rat exocrine pancreas. Cell Tissue Res. 193, 93–105 (1978). https://doi.org/10.1007/BF00221604
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00221604