Entry of newly synthesized GLUT4 into the insulin-responsive storage compartment is GGA dependent

Robert T. Watson, Ahmir H. Khan, Megumi Furukawa, June Chunqiu Hou, Lin Li, Makoto Kanzaki, Shuichi Okada, Konstantin V. Kandror, Jeffrey E. Pessin

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

Following biosynthesis, both GLUT1 and VSV-G proteins appear rapidly (2-3 h) at the plasma membrane, whereas GLUT4 is retained in intracellular membrane compartments and does not display any significant insulin responsiveness until 6-9 h. Surprisingly, the acquisition of insulin responsiveness did not require plasma membrane endocytosis, as expression of a dominant-interfering dynamin mutant (Dyn/K44A) had no effect on the insulin-stimulated GLUT4 translocation. Furthermore, expression of endocytosis-defective GLUT4 mutants or continuous surface labeling with an exofacial specific antibody demonstrated that GLUT4 did not transit the cell surface prior to the acquisition of insulin responsiveness. The expression of a dominant-interfering GGA mutant (VHS-GAT) had no effect on the trafficking of newly synthesized GLUT1 or VSV-G protein to the plasma membrane, but completely blocked the insulin-stimulated translocation of newly synthesized GLUT4. Furthermore, in vitro budding of GLUT4 vesicles but not GLUT1 or the transferrin receptor was inhibited by VHS-GAT. Together, these data demonstrate that following biosynthesis, GLUT4 directly sorts and traffics to the insulin-responsive storage compartment through a specific GGA-sensitive process.

Original languageEnglish (US)
Pages (from-to)2059-2070
Number of pages12
JournalEMBO Journal
Volume23
Issue number10
DOIs
StatePublished - May 19 2004
Externally publishedYes

Fingerprint

Insulin
Cell membranes
Biosynthesis
Cell Membrane
Endocytosis
GTP-Binding Proteins
Dynamins
Intracellular Membranes
Transferrin Receptors
Labeling
Membranes
Antibodies
GAT

Keywords

  • Biosynthesis
  • GGA
  • GLUT4
  • Insulin
  • Trafficking

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Entry of newly synthesized GLUT4 into the insulin-responsive storage compartment is GGA dependent. / Watson, Robert T.; Khan, Ahmir H.; Furukawa, Megumi; Hou, June Chunqiu; Li, Lin; Kanzaki, Makoto; Okada, Shuichi; Kandror, Konstantin V.; Pessin, Jeffrey E.

In: EMBO Journal, Vol. 23, No. 10, 19.05.2004, p. 2059-2070.

Research output: Contribution to journalArticle

Watson, RT, Khan, AH, Furukawa, M, Hou, JC, Li, L, Kanzaki, M, Okada, S, Kandror, KV & Pessin, JE 2004, 'Entry of newly synthesized GLUT4 into the insulin-responsive storage compartment is GGA dependent', EMBO Journal, vol. 23, no. 10, pp. 2059-2070. https://doi.org/10.1038/sj.emboj.7600159
Watson, Robert T. ; Khan, Ahmir H. ; Furukawa, Megumi ; Hou, June Chunqiu ; Li, Lin ; Kanzaki, Makoto ; Okada, Shuichi ; Kandror, Konstantin V. ; Pessin, Jeffrey E. / Entry of newly synthesized GLUT4 into the insulin-responsive storage compartment is GGA dependent. In: EMBO Journal. 2004 ; Vol. 23, No. 10. pp. 2059-2070.
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