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1992 Fiscal Year Final Research Report Summary

Development of insulin sensor using biotechnology Insulin-EGF chimera receptor

Research Project

Project/Area Number 03557033
Research Category

Grant-in-Aid for Developmental Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 内科学一般
Research InstitutionOsaka University

Principal Investigator

AWAMORI Ryuzo  First Dept. Med. Osaka University, 医学部・第一内科, 講師 (00116021)

Co-Investigator(Kenkyū-buntansha) WATARAI Takao  Osaka Univ. Hospital Senior Resident, 医学部・附属病院第一内科, 助手
IWAMA Norimichi  Osaka Univ. Hospital Senior Resident, 医学部・附属病院第一内科, 助手
MORISHIMA Toyohiko  Osaka Univ. Medicine Assistant Teacher, 医学部・第一内科, 助手 (50221635)
YAMAZAKI Yoshimitsu  Osaka Univ. Medicine Assistant Teacher, 医学部・第一内科, 助手 (40201834)
Project Period (FY) 1991 – 1992
KeywordsInsulin biosensor / insulin-EGF chimeric receptor / artophosphorylation
Research Abstract

Cloned human insulin receptor and EGF receptor complementary DNAs were jointed and used in combination with SV40 early promoter transcriptional elements to generate expression construct cording for complete chimeric receptor. This chimeric receptor DNA was inserted into the plasmid pUC12. This plasmid was transfected into COS-7 cells by the calcium phosphate precipitation method. After 48 hr of incubation, insulin-EGF chimeric receptor was partially purified by using WGA-agarose column. We investigated whether this insulin-EGF chimeric receptor was autophosphorylated in a dose-dependent manner.
Analysis by SDS-polyacrylamide gel electrophoresis revealed ^<32>P incorporation into 120KDa and 95KDa proteins, suggesting that 95KDa protein was intrinsic beta subunit of insulin receptor in COS-7 cell and 120KDa protein was beta subunit of transfected insulin-EGF chimeric receptor. Although both insulin receptor and insulin-EGF chimeric receptor were phosphorylated in a dose- dependent manner(insulin concentration, 0-77nM), insulin-EGF chimeric receptor was more highly autophosphorylated than insulin receptor.
These results indicate that this insulin-EGF chimeric receptor can be useful as an insulin biosensor.

  • Research Products

    (7 results)

All Other

All Publications (7 results)

  • [Publications] IWAMA N: "Dephosphorylation of the insulin-receptor partially restores the decreased autophosphorylation in streptozotocin induced diabetic rats." Diabetes Res. 17. 25-32 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] IMANO E: "Rapid Screening method of abnorma insuli-receptor gene expression-Allele-specific oligonucleotide hybridization by using sikent poymorphism." Biochem Biophys Acta in Press. (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] KAJIMOTO Y: "Au AP-l enhancer madiates TPA-induced transcriptional activation of the chioken insulin-like growth factor I gene." Biochemical and Biophysical Research Communis in press.(1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 梶本 佳孝: "IGF-I遺伝子プロモーター構造の解析-GH応答性発現調節におけるC-KinaseおよびAP-1 bindingsiteの重要性" 分子糖尿病学. 3. 7-13 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Iwama N, Watarai T, Kajimoto Y, Yamasaki Y, Yokoyama T, Kawamori R, Kamada T.: "Dephosphorylation of the insulin receptor partially restores the decreased autophosphorylation in streptozotocin induced diabetic rats." Diabetes Res 17. 25-32 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Imono E, Kawamori R, Iwama N, Umayahara Y, Yamasaki Y, Kamada T.: "Rapid screening method of pbnormal insulin-receptor gene expression Allele-specific oligonucleotide hybridization by using sikent poymorphism." Biochem Biophys Acta. (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kajimoto Y, Kawamori R, Umayahara Y, Iwama N, Imano E, Morishima T, Yamasaki Y, Kamada T.: "An AP-1 enhancer madiates TPA-induced transcriptional activation of the chicken insulin-like growth factor I gene." Biochemical and Biophysical Research Communication. (1993)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1994-03-24  

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