Abstract
From rat livers labeled in vivo for 30 min with [35S] cys-met, we have isolated two classes of vesicular carriers operating between the Golgi complex and the basolateral (sinusoidal) plasmalemma. The starting preparation is a Golgi light fraction (GLF) isolated by flotation in a discontinuous sucrose density gradient and processed through immunoisolation on magnetic beads coated with an antibody against the last 11 aa. of the pIgA-R tail. GLF and the ensuing subfractions (bound vs nonbound) were lysed, and the lysates processed through immunoprecipitation with anti-pIgA-R and anti-albumin antibodies followed by radioactivity counting, SDS-PAGE, and fluorography. The recovery of newly synthesized pIgA-R was > 90% and the distribution was 90% vs 10% in the bound vs nonbound subfractions, respectively. Albumin radioactivity was recovered to approximately 80%, with 20% and 80% in bound vs nonbound subfractions, respectively. Other proteins studied were: (a) secretory-apolipoprotein-B, prothrombin, C3 component of the complement, and caeruloplasmin; (b) membrane-transferrin receptor, EGR- receptor, asialoglycoprotein receptor, and the glucose transporter. In all the experiments we have performed, the secretory proteins distributed up to 85% in the nonbound subfraction (large secretory vacuoles), whereas the membrane proteins were segregated up to 95% in the bound subfraction (small vesicular carriers). These results suggest that in hepatocytes, membrane and secretory proteins are transported from the Golgi to the basolateral plasmalemma by separate vesicular carriers as in glandular cells capable of constitutive and regulated secretion.
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- Banting G., Brake B., Braghetta P., Luzio J. P., Stanley K. K. Intracellular targetting signals of polymeric immunoglobulin receptors are highly conserved between species. FEBS Lett. 1989 Aug 28;254(1-2):177–183. doi: 10.1016/0014-5793(89)81034-8. [DOI] [PubMed] [Google Scholar]
- Bartles J. R., Braiterman L. T., Hubbard A. L. Endogenous and exogenous domain markers of the rat hepatocyte plasma membrane. J Cell Biol. 1985 Apr;100(4):1126–1138. doi: 10.1083/jcb.100.4.1126. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boll W., Partin J. S., Katz A. I., Caplan M. J., Jamieson J. D. Distinct pathways for basolateral targeting of membrane and secretory proteins in polarized epithelial cells. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8592–8596. doi: 10.1073/pnas.88.19.8592. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bosshart H., Berger E. G. Biosynthesis and intracellular transport of alpha-2,6-sialyltransferase in rat hepatoma cells. Eur J Biochem. 1992 Sep 1;208(2):341–349. doi: 10.1111/j.1432-1033.1992.tb17192.x. [DOI] [PubMed] [Google Scholar]
- Burgess T. L., Kelly R. B. Constitutive and regulated secretion of proteins. Annu Rev Cell Biol. 1987;3:243–293. doi: 10.1146/annurev.cb.03.110187.001331. [DOI] [PubMed] [Google Scholar]
- Fishman J. B., Fine R. E. A trans Golgi-derived exocytic coated vesicle can contain both newly synthesized cholinesterase and internalized transferrin. Cell. 1987 Jan 16;48(1):157–164. doi: 10.1016/0092-8674(87)90366-7. [DOI] [PubMed] [Google Scholar]
- Fries E., Gustafsson L., Peterson P. A. Four secretory proteins synthesized by hepatocytes are transported from endoplasmic reticulum to Golgi complex at different rates. EMBO J. 1984 Jan;3(1):147–152. doi: 10.1002/j.1460-2075.1984.tb01775.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gottlieb T. A., Beaudry G., Rizzolo L., Colman A., Rindler M., Adesnik M., Sabatini D. D. Secretion of endogenous and exogenous proteins from polarized MDCK cell monolayers. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2100–2104. doi: 10.1073/pnas.83.7.2100. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Green R., Shields D. Somatostatin discriminates between the intracellular pathways of secretory and membrane proteins. J Cell Biol. 1984 Jul;99(1 Pt 1):97–104. doi: 10.1083/jcb.99.1.97. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Howell K. E., Ito A., Palade G. E. Endoplasmic reticulum marker enzymes in Golgi fractions--what does this mean? J Cell Biol. 1978 Nov;79(2 Pt 1):581–589. doi: 10.1083/jcb.79.2.581. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Howell K. E., Palade G. E. Hepatic Golgi fractions resolved into membrane and content subfractions. J Cell Biol. 1982 Mar;92(3):822–832. doi: 10.1083/jcb.92.3.822. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Howell K. E., Palade G. E. Heterogeneity of lipoprotein particles in hepatic Golgi fractions. J Cell Biol. 1982 Mar;92(3):833–845. doi: 10.1083/jcb.92.3.833. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ladinsky M. S., Howell K. E. The trans-Golgi network can be dissected structurally and functionally from the cisternae of the Golgi complex by brefeldin A. Eur J Cell Biol. 1992 Oct;59(1):92–105. [PubMed] [Google Scholar]
- Larkin J. M., Sztul E. S., Palade G. E. Phosphorylation of the rat hepatic polymeric IgA receptor. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4759–4763. doi: 10.1073/pnas.83.13.4759. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McCaffrey G., Jamieson J. C. Evidence for the role of a cathepsin D-like activity in the release of Gal beta 1-4GlcNAc alpha 2-6sialyltransferase from rat and mouse liver in whole-cell systems. Comp Biochem Physiol B. 1993 Jan;104(1):91–94. doi: 10.1016/0305-0491(93)90342-3. [DOI] [PubMed] [Google Scholar]
- Musil L. S., Baenziger J. U. Cleavage of membrane secretory component to soluble secretory component occurs on the cell surface of rat hepatocyte monolayers. J Cell Biol. 1987 Jun;104(6):1725–1733. doi: 10.1083/jcb.104.6.1725. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Palade G. Intracellular aspects of the process of protein synthesis. Science. 1975 Aug 1;189(4200):347–358. doi: 10.1126/science.1096303. [DOI] [PubMed] [Google Scholar]
- Parczyk K., Haase W., Kondor-Koch C. Microtubules are involved in the secretion of proteins at the apical cell surface of the polarized epithelial cell, Madin-Darby canine kidney. J Biol Chem. 1989 Oct 5;264(28):16837–16846. [PubMed] [Google Scholar]
- Redman C. M., Banerjee D., Howell K., Palade G. E. Colchicine inhibition of plasma protein release from rat hepatocytes. J Cell Biol. 1975 Jul;66(1):42–59. doi: 10.1083/jcb.66.1.42. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Salamero J., Sztul E. S., Howell K. E. Exocytic transport vesicles generated in vitro from the trans-Golgi network carry secretory and plasma membrane proteins. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7717–7721. doi: 10.1073/pnas.87.19.7717. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Saucan L., Palade G. E. Differential colchicine effects on the transport of membrane and secretory proteins in rat hepatocytes in vivo: bipolar secretion of albumin. Hepatology. 1992 Apr;15(4):714–721. doi: 10.1002/hep.1840150427. [DOI] [PubMed] [Google Scholar]
- Schnitzer J. E., Carley W. W., Palade G. E. Specific albumin binding to microvascular endothelium in culture. Am J Physiol. 1988 Mar;254(3 Pt 2):H425–H437. doi: 10.1152/ajpheart.1988.254.3.H425. [DOI] [PubMed] [Google Scholar]
- Stein B. S., Sussman H. H. Demonstration of two distinct transferrin receptor recycling pathways and transferrin-independent receptor internalization in K562 cells. J Biol Chem. 1986 Aug 5;261(22):10319–10331. [PubMed] [Google Scholar]
- Stein O., Sanger L., Stein Y. Colchicine-induced inhibition of lipoprotein and protein secretion into the serum and lack of interference with secretion of biliary phospholipids and cholesterol by rat liver in vivo. J Cell Biol. 1974 Jul;62(1):90–103. doi: 10.1083/jcb.62.1.90. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Strous G. J. Golgi and secreted galactosyltransferase. CRC Crit Rev Biochem. 1986;21(2):119–151. doi: 10.3109/10409238609113610. [DOI] [PubMed] [Google Scholar]
- Sztul E. S., Howell K. E., Palade G. E. Biogenesis of the polymeric IgA receptor in rat hepatocytes. II. Localization of its intracellular forms by cell fractionation studies. J Cell Biol. 1985 Apr;100(4):1255–1261. doi: 10.1083/jcb.100.4.1255. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sztul E., Kaplin A., Saucan L., Palade G. Protein traffic between distinct plasma membrane domains: isolation and characterization of vesicular carriers involved in transcytosis. Cell. 1991 Jan 11;64(1):81–89. doi: 10.1016/0092-8674(91)90210-p. [DOI] [PubMed] [Google Scholar]
- Velasco A., Hendricks L., Moremen K. W., Tulsiani D. R., Touster O., Farquhar M. G. Cell type-dependent variations in the subcellular distribution of alpha-mannosidase I and II. J Cell Biol. 1993 Jul;122(1):39–51. doi: 10.1083/jcb.122.1.39. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Curtis I., Simons K. Isolation of exocytic carrier vesicles from BHK cells. Cell. 1989 Aug 25;58(4):719–727. doi: 10.1016/0092-8674(89)90106-2. [DOI] [PubMed] [Google Scholar]