| Project/Area Number |
63870046
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Research Field |
内分泌・代謝学
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| Research Institution | University of Tokyo |
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Principal Investigator |
OKA Yoshitomo Faculty of Medicine, University of Tokyo Assistant Professor, 医学部(病), 助手 (70175256)
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| Co-Investigator(Kenkyū-buntansha) |
AKANUMA Yasuo Institute for Diabetes Care and Research, Asahi Life Foundation Director, 所長
ASANO Tomoichiro Faculty of Medicine, University of Tokyo Research Associate, 医学部(病), 医員
SHIBASAKI Yoshikazu Faculty of Medicine, University of Tokyo Assistant Professor, 医学部(病), 助手 (80196419)
春日 雅人 東京大学, 医学部(病), 講師 (50161047)
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| Project Period (FY) |
1988 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥13,000,000 (Direct Cost: ¥13,000,000)
Fiscal Year 1990: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1989: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1988: ¥10,500,000 (Direct Cost: ¥10,500,000)
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| Keywords | Radioimmunoassay / Glucose transporter / Synthetic Peptides / ラジオイムノアッセイ |
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| Research Abstract |
A radioimmunoassay (RIA) for the GLUT1 glucose transporter has been developed by using a synthesized peptide based upon the sequence of the cDNA for GLUTI1. A peptide corresponding to the carboxy-terminal domain of the GLUT1 glucose transporter (ThrProGluGluLeuPheHisProLeuGlyAlaAspSerGlnVal) was synthesized and conjugated to keyhole limpet hemocyanin through the amino-terminus of the peptide. An antibody was raised against this complex and affinity-purified using the immobilized peptide. A second peptide with tyrosine residue added to amino-terminus of the above peptide was synthesized and used as a standard and also iodinated for preparation of the radioactive ligand. The assay is highly reproducible, sensitive, and specific for the carboxy-terminal domain of the GLUT1 glucose transporter. It has no cross-reactivity with the other glucose transporter isoforms, GLUT2 and GLUT4. Furthermore, the results obtained with this RIA on the number of glucose transporters in human erythrocytes are in a fairly good agreement with the previous reports based on cytochalasin B binding, suggesting that this RIA is able to quantify the number of glucose transporters. The assay is completed within 4h and can be used for simultaneous measurement of GLUT1 in many samples. In addition, it can be applied to the measurement of GLUT1 in several types of tissue.
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