{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,7]],"date-time":"2026-02-07T22:43:08Z","timestamp":1770504188566,"version":"3.49.0"},"reference-count":74,"publisher":"Rockefeller University Press","issue":"3","content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[1998,2,9]]},"abstract":"<jats:p>The trafficking of GLUT4, a facilitative glucose transporter, is examined in transfected CHO cells. In previous work, we expressed GLUT4 in neuroendocrine cells and fibroblasts and found that it was targeted to a population of small vesicles slightly larger than synaptic vesicles (Herman, G.A, F. Bonzelius, A.M. Cieutat, and R.B. Kelly. 1994. Proc. Natl. Acad. Sci. USA. 91: 12750\u201312754.). In this study, we demonstrate that at 37\u00b0C, GLUT4-containing small vesicles (GSVs) are detected after cell surface radiolabeling of GLUT4 whereas uptake of radioiodinated human transferrin does not show appreciable accumulation within these small vesicles. Immunofluorescence microscopy experiments show that at 37\u00b0C, cell surface\u2013labeled GLUT4 as well as transferrin is internalized into peripheral and perinuclear structures. At 15\u00b0C, endocytosis of GLUT4 continues to occur at a slowed rate, but whereas fluorescently labeled GLUT4 is seen to accumulate within large peripheral endosomes, no perinuclear structures are labeled, and no radiolabeled GSVs are detectable. Shifting cells to 37\u00b0C after accumulating labeled GLUT4 at 15\u00b0C results in the reappearance of GLUT4 in perinuclear structures and GSV reformation. Cytosol acidification or treatment with hypertonic media containing sucrose prevents the exit of GLUT4 from peripheral endosomes as well as GSV formation, suggesting that coat proteins may be involved in the endocytic trafficking of GLUT4. In contrast, at 15\u00b0C, transferrin continues to traffic to perinuclear structures and overall labels structures similar in distribution to those observed at 37\u00b0C. Furthermore, treatment with hypertonic media has no apparent effect on transferrin trafficking from peripheral endosomes. Double-labeling experiments after the internalization of both transferrin and surface-labeled GLUT4 show that GLUT4 accumulates within peripheral compartments that exclude the transferrin receptor (TfR) at both 15\u00b0 and 37\u00b0C. Thus, GLUT4 is sorted differently from the transferrin receptor as evidenced by the targeting of each protein to distinct early endosomal compartments and by the formation of GSVs. These results suggest that the sorting of GLUT4 from TfR may occur primarily at the level of the plasma membrane into distinct endosomes and that the organization of the endocytic system in CHO cells more closely resembles that of neuroendocrine cells than previously appreciated.<\/jats:p>","DOI":"10.1083\/jcb.140.3.565","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T16:47:50Z","timestamp":1027702070000},"page":"565-575","update-policy":"https:\/\/doi.org\/10.1083\/jcb.crossmarkpolicy","source":"Crossref","is-referenced-by-count":50,"title":["GLUT4 and Transferrin Receptor Are Differentially Sorted Along the Endocytic Pathway in CHO Cells"],"prefix":"10.1083","volume":"140","author":[{"given":"Maria L.","family":"Wei","sequence":"first","affiliation":[{"name":"*Department of Biochemistry and Biophysics, \u2021Department of Dermatology, \u2016The Hormone Research Institute; \u00a7Department of Pediatrics, Division of Gastroenterology and Nutrition, University of California, San Francisco, California 94143"},{"name":"*Department of Biochemistry and Biophysics, \u2021Department of Dermatology, \u2016The Hormone Research Institute; \u00a7Department of Pediatrics, Division of Gastroenterology and Nutrition, University of California, San Francisco, California 94143"},{"name":"*Department of Biochemistry and Biophysics, \u2021Department of Dermatology, \u2016The Hormone Research Institute; \u00a7Department of Pediatrics, Division of Gastroenterology and Nutrition, University of California, San Francisco, California 94143"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Frank","family":"Bonzelius","sequence":"additional","affiliation":[{"name":"*Department of Biochemistry and Biophysics, \u2021Department of Dermatology, \u2016The Hormone Research Institute; \u00a7Department of Pediatrics, Division of Gastroenterology and Nutrition, University of California, San Francisco, California 94143"},{"name":"*Department of Biochemistry and Biophysics, \u2021Department of Dermatology, \u2016The Hormone Research Institute; \u00a7Department of Pediatrics, Division of Gastroenterology and Nutrition, University of California, San Francisco, California 94143"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rebecca M.","family":"Scully","sequence":"additional","affiliation":[{"name":"*Department of Biochemistry and Biophysics, \u2021Department of Dermatology, \u2016The Hormone Research Institute; \u00a7Department of Pediatrics, Division of Gastroenterology and Nutrition, University of California, San Francisco, California 94143"},{"name":"*Department of Biochemistry and Biophysics, \u2021Department of Dermatology, \u2016The Hormone Research Institute; \u00a7Department of Pediatrics, Division of Gastroenterology and Nutrition, University of California, San Francisco, California 94143"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Regis B.","family":"Kelly","sequence":"additional","affiliation":[{"name":"*Department of Biochemistry and Biophysics, \u2021Department of Dermatology, \u2016The Hormone Research Institute; \u00a7Department of Pediatrics, Division of Gastroenterology and Nutrition, University of California, San Francisco, California 94143"},{"name":"*Department of Biochemistry and Biophysics, \u2021Department of Dermatology, \u2016The Hormone Research Institute; \u00a7Department of Pediatrics, Division of Gastroenterology and Nutrition, University of California, San Francisco, California 94143"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gary A.","family":"Herman","sequence":"additional","affiliation":[{"name":"*Department of Biochemistry and Biophysics, \u2021Department of Dermatology, \u2016The Hormone Research Institute; \u00a7Department of Pediatrics, Division of Gastroenterology and Nutrition, University of California, San Francisco, California 94143"},{"name":"*Department of Biochemistry and Biophysics, \u2021Department of Dermatology, \u2016The Hormone Research Institute; \u00a7Department of Pediatrics, Division of Gastroenterology and Nutrition, University of California, San Francisco, California 94143"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"291","published-online":{"date-parts":[[1998,2,9]]},"reference":[{"key":"2023072207405274300_B1","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1083\/jcb.133.1.29","article-title":"An endosomal \u03b2 COP is involved in the pH-dependent formation of transport vesicles destined for late endosomes","volume":"133","author":"Aniento","year":"1996","journal-title":"J Cell Biol"},{"key":"2023072207405274300_B2","doi-asserted-by":"crossref","first-page":"16180","DOI":"10.1016\/S0021-9258(18)66695-5","article-title":"Isolation of vesicles containing insulin- responsive, intracellular glucose transporters from 3T3-L1 adipocytes","volume":"261","author":"Biber","year":"1986","journal-title":"J Biol Chem"},{"key":"2023072207405274300_B3","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1083\/jcb.106.1.69","article-title":"Insulin-induced translocation of glucose transporters from post-Golgi compartments to the plasma membrane of 3T3-L1 adipocytes","volume":"106","author":"Blok","year":"1988","journal-title":"J Cell Biol"},{"key":"2023072207405274300_B4","doi-asserted-by":"crossref","first-page":"1603","DOI":"10.1083\/jcb.127.6.1603","article-title":"The polymeric immunoglobulin receptor accumulates in specialized endosomes but not synaptic vesicles within the neurites of transfected neuroendocrine PC12 cells","volume":"127","author":"Bonzelius","year":"1994","journal-title":"J Cell Biol"},{"key":"2023072207405274300_B5","doi-asserted-by":"crossref","first-page":"1177","DOI":"10.1083\/jcb.133.6.1177","article-title":"A biosynthetic regulated secretory pathway in constitutive secretory cells","volume":"133","author":"Chavez","year":"1996","journal-title":"J Cell Biol"},{"key":"2023072207405274300_B6","doi-asserted-by":"crossref","first-page":"1693","DOI":"10.1083\/jcb.110.5.1693","article-title":"Biogenesis of synaptic vesicle-like structures in a pheochromocytoma cell line PC-12","volume":"110","author":"Clift-O'Grady","year":"1990","journal-title":"J Cell Biol"},{"key":"2023072207405274300_B7","doi-asserted-by":"crossref","first-page":"3787","DOI":"10.1002\/j.1460-2075.1996.tb00752.x","article-title":"Ca2+triggers massive exocytosis in Chinese hamster ovary cells","volume":"15","author":"Coorssen","year":"1996","journal-title":"EMBO (Eur Mol Biol Organ) J"},{"key":"2023072207405274300_B8","doi-asserted-by":"crossref","first-page":"19491","DOI":"10.1016\/S0021-9258(19)36542-1","article-title":"Insulin and okadaic acid induce rab4 redistribution in adipocytes","volume":"268","author":"Cormont","year":"1993","journal-title":"J Biol Chem"},{"key":"2023072207405274300_B9","doi-asserted-by":"crossref","first-page":"3408","DOI":"10.1210\/endo.137.8.8754768","article-title":"Insulin induces a change in rab5 subcellular localization in adipocytes independently of phosphatidylinositol 3-kinase activation","volume":"137","author":"Cormont","year":"1996","journal-title":"Endocrinology"},{"key":"2023072207405274300_B10","doi-asserted-by":"crossref","first-page":"979","DOI":"10.1083\/jcb.126.4.979","article-title":"A double leucine within the GLUT4 glucose transporter COOH-terminal domain functions as an endocytosis signal","volume":"126","author":"Corvera","year":"1994","journal-title":"J Cell Biol"},{"key":"2023072207405274300_B11","doi-asserted-by":"crossref","first-page":"4758","DOI":"10.1016\/S0021-9258(19)85561-8","article-title":"Potential mechanism of insulin action on glucose transport in the isolated rat adipose cell. 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