The functional analysis of a novel adaptor protein binding to GLUT4
| Project/Area Number |
15590939
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Metabolomics
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| Research Institution | Yamaguchi University |
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Principal Investigator |
OKUYA Shigeru Yamaguchi University, Graduate School of Medicine, Associate Professor, 大学院・医学研究科, 助教授 (20214083)
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| Co-Investigator(Kenkyū-buntansha) |
EMOTO Masahiro Yamaguchi University, University Hospital, Research Associate, 大学院・医学部附属病院, 助手 (50294640)
UEDA Kohei Yamaguchi University, Health Service Center, Research Associate, 大学教育機構保健管理センター, 助手 (50325221)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2004: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2003: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | glucose transporter / GLUT4 / translocation / recycling / adaptor protein / glucose uptake / insulin / adipocyte |
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| Research Abstract |
Purpose
It has been supposed that a protein binding to GLUT4 could play an important role during the recycling of GLUT4 vesicle and insulin-induced GLUT4 translocation. We assumed the existence of GLUT4 "adaptor protein" in adipocytes, and we have identified a novel protein "p61" that interacts with GLUT4, and analyzed its function.
Method
Using the yeast two-hybrid system, we screened from rat adipocyte cDNA library, and have cloned p61 as an interesting protein that interacts with GLUT4. The intracellular localization of p61 in L6-GLUT4myc myotubes, stably expressing myc-tagged GLUT4, was examined by the immunohistochemistry using the myc tag antibody. The interaction of GLUT4 and p61 was confirmed by the yeast two-hybrid system, and also the immunoprecipitation. After overexpression of p61 in 3T3-L1 adipocytes, the GLUT4 translocation and the insulin-induced glucose uptake were examined.
Results
The expression of p61 in adipocytes was confirmed at the mRNA level using Northern blotting. The p61 consists of 540 amino acids with the ANK (ankyrin) structure thought to be a protein-protein interaction motif. The p61 existed in perinuclear region in the L6-GLUT4myc myotubes as a result of the immunohistochemistry. Moreover, it was confirmed that the interaction of p61 and GLUT4 by yeast two-hybrid system and the immunoprecipitation. In addition, overexpression of p61 in 3T3-L1 adipocytes using adenoviral system, increased the amount of GLUT4 in membrane fraction after insulin stimulation, and also significantly facilitated insulin-induced glucose uptake.
Conclusion
It is suggested that a novel ANK structure protein p61 facilitates GLUT4 translocation, and then activates insulin-induced glucose uptake.
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Report
(3 results)
Research Products
(4 results)
