Abstract
Receptor-mediated endocytosis and intracellular routing of native ricin, and of ricin conjugated to colloidal gold (Ri-Au) and to horseradish peroxidase (Ri-HRP), have been studied in cultured MCF-7 and Vero cells by electron microscopical techniques including serial section analysis. Both native ricin, as demonstrated by immunoperoxidase cytochemistry, and the ricin conjugates were internalized via a common coated pit-coated vesicle pathway to reach vacuolar and tubulo-vesicular portions of the endosomal system. In addition, native ricin and a purified monovalent fraction of Ri-HRP reached distinct Golgi cisterns, whereas Ri-Au and polyvalent Ri-HRP did not. The results delineate intracellular routing of native ricin and compare it with the routing of different ricin conjugates. Moreover, our study shows that conjugates of a particular ligand (ricin) and various probes (e.g., gold and peroxidase), may be handled differently by cells. Sorting apparently takes place in the endosomal system, allowing some but not other molecules to reach Golgi elements. This sorting seems to depend on the valency of the ricin conjugate.
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- Anderson R. G., Brown M. S., Beisiegel U., Goldstein J. L. Surface distribution and recycling of the low density lipoprotein receptor as visualized with antireceptor antibodies. J Cell Biol. 1982 Jun;93(3):523–531. doi: 10.1083/jcb.93.3.523. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Anderson R. G., Falck J. R., Goldstein J. L., Brown M. S. Visualization of acidic organelles in intact cells by electron microscopy. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4838–4842. doi: 10.1073/pnas.81.15.4838. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Anderson R. G., Pathak R. K. Vesicles and cisternae in the trans Golgi apparatus of human fibroblasts are acidic compartments. Cell. 1985 Mar;40(3):635–643. doi: 10.1016/0092-8674(85)90212-0. [DOI] [PubMed] [Google Scholar]
- Bjorn M. J., Ring D., Frankel A. Evaluation of monoclonal antibodies for the development of breast cancer immunotoxins. Cancer Res. 1985 Mar;45(3):1214–1221. [PubMed] [Google Scholar]
- CALLAWAY E., BAND R. I. Some psychopharmacological effects of atropine; preliminary investigation of broadened attention. AMA Arch Neurol Psychiatry. 1958 Jan;79(1):91–102. doi: 10.1001/archneurpsyc.1958.02340010109010. [DOI] [PubMed] [Google Scholar]
- Carlsson J., Drevin H., Axén R. Protein thiolation and reversible protein-protein conjugation. N-Succinimidyl 3-(2-pyridyldithio)propionate, a new heterobifunctional reagent. Biochem J. 1978 Sep 1;173(3):723–737. doi: 10.1042/bj1730723. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carrière D., Casellas P., Richer G., Gros P., Jansen F. K. Endocytosis of an antibody ricin A-chain conjugate (immuno-A-toxin) adsorbed on colloidal gold. Effects of ammonium chloride and monensin. Exp Cell Res. 1985 Feb;156(2):327–340. [PubMed] [Google Scholar]
- Christensen E. I. Rapid membrane recycling in renal proximal tubule cells. Eur J Cell Biol. 1982 Nov;29(1):43–49. [PubMed] [Google Scholar]
- Farquhar M. G., Palade G. E. The Golgi apparatus (complex)-(1954-1981)-from artifact to center stage. J Cell Biol. 1981 Dec;91(3 Pt 2):77s–103s. doi: 10.1083/jcb.91.3.77s. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Farquhar M. G. Recovery of surface membrane in anterior pituitary cells. Variations in traffic detected with anionic and cationic ferritin. J Cell Biol. 1978 Jun;77(3):R35–R42. doi: 10.1083/jcb.77.3.r35. [DOI] [PMC free article] [PubMed] [Google Scholar]
- FitzGerald D., Morris R. E., Saelinger C. B. Receptor-mediated internalization of Pseudomonas toxin by mouse fibroblasts. Cell. 1980 Oct;21(3):867–873. doi: 10.1016/0092-8674(80)90450-x. [DOI] [PubMed] [Google Scholar]
- Geuze H. J., Slot J. W., Strous G. J., Lodish H. F., Schwartz A. L. Intracellular site of asialoglycoprotein receptor-ligand uncoupling: double-label immunoelectron microscopy during receptor-mediated endocytosis. Cell. 1983 Jan;32(1):277–287. doi: 10.1016/0092-8674(83)90518-4. [DOI] [PubMed] [Google Scholar]
- Geuze H. J., Slot J. W., Strous G. J., Peppard J., von Figura K., Hasilik A., Schwartz A. L. Intracellular receptor sorting during endocytosis: comparative immunoelectron microscopy of multiple receptors in rat liver. Cell. 1984 May;37(1):195–204. doi: 10.1016/0092-8674(84)90315-5. [DOI] [PubMed] [Google Scholar]
- Gonatas J., Stieber A., Olsnes S., Gonatas N. K. Pathways involved in fluid phase and adsorptive endocytosis in neuroblastoma. J Cell Biol. 1980 Dec;87(3 Pt 1):579–588. doi: 10.1083/jcb.87.3.579. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gonatas N. K., Stieber A., Hickey W. F., Herbert S. H., Gonatas J. O. Endosomes and Golgi vesicles in adsorptive and fluid phase endocytosis. J Cell Biol. 1984 Oct;99(4 Pt 1):1379–1390. doi: 10.1083/jcb.99.4.1379. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gonnella P. A., Neutra M. R. Membrane-bound and fluid-phase macromolecules enter separate prelysosomal compartments in absorptive cells of suckling rat ileum. J Cell Biol. 1984 Sep;99(3):909–917. doi: 10.1083/jcb.99.3.909. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hopkins C. R. Intracellular routing of transferrin and transferrin receptors in epidermoid carcinoma A431 cells. Cell. 1983 Nov;35(1):321–330. doi: 10.1016/0092-8674(83)90235-0. [DOI] [PubMed] [Google Scholar]
- Hopkins C. R., Trowbridge I. S. Internalization and processing of transferrin and the transferrin receptor in human carcinoma A431 cells. J Cell Biol. 1983 Aug;97(2):508–521. doi: 10.1083/jcb.97.2.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Joseph K. C., Stieber A., Gonatas N. K. Endocytosis of cholera toxin in GERL-like structures of murine neuroblastoma cells pretreated with GM1 ganglioside. Cholera toxin internalization into Neuroblastoma GERL. J Cell Biol. 1979 Jun;81(3):543–554. doi: 10.1083/jcb.81.3.543. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Louvard D., Reggio H., Warren G. Antibodies to the Golgi complex and the rough endoplasmic reticulum. J Cell Biol. 1982 Jan;92(1):92–107. doi: 10.1083/jcb.92.1.92. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Madshus I. H., Olsnes S., Sandvig K. Mechanism of entry into the cytosol of poliovirus type 1: requirement for low pH. J Cell Biol. 1984 Apr;98(4):1194–1200. doi: 10.1083/jcb.98.4.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marsh M. The entry of enveloped viruses into cells by endocytosis. Biochem J. 1984 Feb 15;218(1):1–10. doi: 10.1042/bj2180001. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mellman I., Plutner H. Internalization and degradation of macrophage Fc receptors bound to polyvalent immune complexes. J Cell Biol. 1984 Apr;98(4):1170–1177. doi: 10.1083/jcb.98.4.1170. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mellman I., Plutner H., Ukkonen P. Internalization and rapid recycling of macrophage Fc receptors tagged with monovalent antireceptor antibody: possible role of a prelysosomal compartment. J Cell Biol. 1984 Apr;98(4):1163–1169. doi: 10.1083/jcb.98.4.1163. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Montesano R., Roth J., Robert A., Orci L. Non-coated membrane invaginations are involved in binding and internalization of cholera and tetanus toxins. Nature. 1982 Apr 15;296(5858):651–653. doi: 10.1038/296651a0. [DOI] [PubMed] [Google Scholar]
- Nicolson G. L., Lacorbiere M., Hunter T. R. Mechanism of cell entry and toxicity of an affinity- purified lectin from Ricinus communis and its differential effects on normal and virus-transformed fibroblasts. Cancer Res. 1975 Jan;35(1):144–155. [PubMed] [Google Scholar]
- Nicolson G. L., Smith J. R., Hyman R. Dynamics of toxin and lectin receptors on a lymphoma cell line and its toxin-resistant variant using ferritin-conjugated, 125I-labeled ligand. J Cell Biol. 1978 Aug;78(2):565–576. doi: 10.1083/jcb.78.2.565. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ottosen P. D., Courtoy P. J., Farquhar M. G. Pathways followed by membrane recovered from the surface of plasma cells and myeloma cells. J Exp Med. 1980 Jul 1;152(1):1–19. doi: 10.1084/jem.152.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Petersen O. W., van Deurs B. Serial-section analysis of coated pits and vesicles involved in adsorptive pinocytosis in cultured fibroblasts. J Cell Biol. 1983 Jan;96(1):277–281. doi: 10.1083/jcb.96.1.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sandvig K., Olsnes S. Diphtheria toxin entry into cells is facilitated by low pH. J Cell Biol. 1980 Dec;87(3 Pt 1):828–832. doi: 10.1083/jcb.87.3.828. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sandvig K., Olsnes S. Effect of temperature on the uptake, excretion and degradation of abrin and ricin by HeLa cells. Exp Cell Res. 1979 Jun;121(1):15–25. doi: 10.1016/0014-4827(79)90439-7. [DOI] [PubMed] [Google Scholar]
- Sandvig K., Olsnes S. Entry of the toxic proteins abrin, modeccin, ricin, and diphtheria toxin into cells. I. Requirement for calcium. J Biol Chem. 1982 Jul 10;257(13):7495–7503. [PubMed] [Google Scholar]
- Sandvig K., Olsnes S. Entry of the toxic proteins abrin, modeccin, ricin, and diphtheria toxin into cells. II. Effect of pH, metabolic inhibitors, and ionophores and evidence for toxin penetration from endocytotic vesicles. J Biol Chem. 1982 Jul 10;257(13):7504–7513. [PubMed] [Google Scholar]
- Sandvig K., Olsnes S. Receptor-mediated entry of protein toxins into cells. Acta Histochem Suppl. 1984;29:79–94. [PubMed] [Google Scholar]
- Sandvig K., Sundan A., Olsnes S. Evidence that diphtheria toxin and modeccin enter the cytosol from different vesicular compartments. J Cell Biol. 1984 Mar;98(3):963–970. doi: 10.1083/jcb.98.3.963. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schwartz A. L., Strous G. J., Slot J. W., Geuze H. J. Immunoelectron microscopic localization of acidic intracellular compartments in hepatoma cells. EMBO J. 1985 Apr;4(4):899–904. doi: 10.1002/j.1460-2075.1985.tb03716.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Soule H. D., Vazguez J., Long A., Albert S., Brennan M. A human cell line from a pleural effusion derived from a breast carcinoma. J Natl Cancer Inst. 1973 Nov;51(5):1409–1416. doi: 10.1093/jnci/51.5.1409. [DOI] [PubMed] [Google Scholar]
- Steinman R. M., Mellman I. S., Muller W. A., Cohn Z. A. Endocytosis and the recycling of plasma membrane. J Cell Biol. 1983 Jan;96(1):1–27. doi: 10.1083/jcb.96.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thyberg J., Nilsson J., Hellgren D. Recirculation of cationized ferritin in cultured mouse peritoneal macrophages. Electron microscopic and cytochemical studies with double-labeling technique. Eur J Cell Biol. 1980 Dec;23(1):85–94. [PubMed] [Google Scholar]
- Tougard C., Picart R., Tixier-Vidal A. Electron-microscopic cytochemical studies on the secretory process in rat prolactin cells in primary culture. Am J Anat. 1980 Aug;158(4):471–490. doi: 10.1002/aja.1001580409. [DOI] [PubMed] [Google Scholar]
- Van Deurs B., Nilausen K. Pinocytosis in mouse L-fibroblasts: ultrastructural evidence for a direct membrane shuttle between the plasma membrane and the lysosomal compartment. J Cell Biol. 1982 Aug;94(2):279–286. doi: 10.1083/jcb.94.2.279. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vitetta E. S., Krolick K. A., Miyama-Inaba M., Cushley W., Uhr J. W. Immunotoxins: a new approach to cancer therapy. Science. 1983 Feb 11;219(4585):644–650. doi: 10.1126/science.6218613. [DOI] [PubMed] [Google Scholar]
- Willingham M. C., Hanover J. A., Dickson R. B., Pastan I. Morphologic characterization of the pathway of transferrin endocytosis and recycling in human KB cells. Proc Natl Acad Sci U S A. 1984 Jan;81(1):175–179. doi: 10.1073/pnas.81.1.175. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Willingham M. C., Pastan I. H. Transit of epidermal growth factor through coated pits of the Golgi system. J Cell Biol. 1982 Jul;94(1):207–212. doi: 10.1083/jcb.94.1.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Willingham M. C., Pastan I. Endocytosis and exocytosis: current concepts of vesicle traffic in animal cells. Int Rev Cytol. 1984;92:51–92. doi: 10.1016/s0074-7696(08)61324-8. [DOI] [PubMed] [Google Scholar]
- Yamashiro D. J., Tycko B., Fluss S. R., Maxfield F. R. Segregation of transferrin to a mildly acidic (pH 6.5) para-Golgi compartment in the recycling pathway. Cell. 1984 Jul;37(3):789–800. doi: 10.1016/0092-8674(84)90414-8. [DOI] [PubMed] [Google Scholar]
- Yokoyama M., Nishiyama F., Kawai N., Hirano H. The staining of Golgi membranes with Ricinus communis agglutinin-horseradish peroxidase conjugate in mice tissue cells. Exp Cell Res. 1980 Jan;125(1):47–53. doi: 10.1016/0014-4827(80)90187-1. [DOI] [PubMed] [Google Scholar]
- Zentz C., Frénoy J. P., Bourrillon R. Binding of galactose and lactose to ricin. Equilibrium studies. Biochim Biophys Acta. 1978 Sep 26;536(1):18–26. doi: 10.1016/0005-2795(78)90047-8. [DOI] [PubMed] [Google Scholar]
- van Deurs B., Christensen E. I. Endocytosis in kidney proximal tubule cells and cultured fibroblasts: a review of the structural aspects of membrane recycling between the plasma membrane and endocytic vacuoles. Eur J Cell Biol. 1984 Jan;33(1):163–173. [PubMed] [Google Scholar]
- van Deurs B., Pedersen L. R., Sundan A., Olsnes S., Sandvig K. Receptor-mediated endocytosis of a ricin-colloidal gold conjugate in vero cells. Intracellular routing to vacuolar and tubulo-vesicular portions of the endosomal system. Exp Cell Res. 1985 Aug;159(2):287–304. doi: 10.1016/s0014-4827(85)80003-3. [DOI] [PubMed] [Google Scholar]
- van Deurs B., Petersen O. W., Bundgaard M. Identification of free coated pinocytic vesicles in Swiss 3T3 cells. EMBO J. 1984 Sep;3(9):1959–1964. doi: 10.1002/j.1460-2075.1984.tb02076.x. [DOI] [PMC free article] [PubMed] [Google Scholar]