Analysis of molecular basis of insulin receptor and its function with use of synthetic peptides
| Project/Area Number |
62480257
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| Research Category |
Grant-in-Aid for General 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 | Osaka University |
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Principal Investigator |
YANAIHARA Chizuko Osaka University Hospital, Professor, 医学部附属病院, 教授 (00046252)
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| Project Period (FY) |
1987 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥5,100,000 (Direct Cost: ¥5,100,000)
Fiscal Year 1989: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1988: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1987: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | Insulin receptor / Synthetic peptide / Anti-synthetic peptide antiserum / Insulin receptor binding / Immunostaining of insulin receptor / Radioimmunoassay specific for insulin receptor / Insulin receptor-specific antibody / Insulin-binding inhibition / 抗合成ペプチド抗体 / インスリン受容体RIA |
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| Research Abstract |
1. Chemical synthesis of peptides related to human placental insulin receptor protein (HIRP): Based on the predicted amino acid sequence of HIRP(1-1355), chemical synthesis of polypeptides related to HIRP was accomplished by a solid phase technique. Among them, ten peptides were successfully used as antigens for production of HIRP-specific antisera.
2. Production of anti-synthetic peptide antisera: Using the HIRP-related synthetic peptides as antigens, seventeen kinds of region-specific antisera were produced in rabbits, among which ten antisera were used in the present study.
3. Development of HIRP-specific radioimmunoassays: Seven kinds of radioimmunoassay systems specific for HIRP were developed with use of synthetic peptides and anti-synthetic peptide antisera.
4. Immunohistochemical study of insulin receptor: The present anti-synthetic peptide antisera specific for HIRP immunostained the membrane regions of syncytiotrophoblast as well as Hofbauer cells in human placenta.
5. Biochemica
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l study of insulin receptor with use of anti-synthetic peptide antidera and radioimmunoassays: (1) Two distinct insulinbinding components were demonstrated in solubilized human placental membranes and the characteristic effects of pH and salt concentration on the binding abilities of the components were disclosed. (2) The presence of insulin receptor in human hepatoma PLC/PRF/5 cells was demonstrated and the insulin binding sites were immunochemically analyzed. (3) An amino-terminal portion of a-subunit of human placental HIRP was shown to participate in interaction of the receptor with insulin. (4) In human placental insulin receptor, a carboxyl-terminal portion of alpha-subunit and a portion of beta-subunit located very close to cell membrane were both suggested to be on the surface of the receptor molecules in either alphaB or alpha_2B_2 form. (5) The presennthetic peptide antisera and radioimmunoassays were also effectively applied for immunochemical analysis of insulin-binding sites of human monocute. Less
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Report
(4 results)
Research Products
(35 results)
