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Study on Specificity of Hormone Action of Growth Hormone/Prolactin Gene Family

Research Project

Project/Area Number 01580188
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 代謝生物化学
Research InstitutionMie University

Principal Investigator

TANAKA Minoru  Mie University, School of Medicine, Lecturer, 医学部, 講師 (90024736)

Co-Investigator(Kenkyū-buntansha) USHIRO Hiroshi  Mie University, School of Medicine, Lecturer, 医学部, 講師 (10151854)
NAKASHIMA Kunio  Mie University, School of Medicine, Professor, 医学部, 教授 (40022800)
Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1990: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1989: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsHuman Growth Hormone / Human Prolactin / Flounder Growth Hormone / Bovine Prolactin-like Hormone / cDNA Cloning / Biological Activity / 成長ホルモン / プロラクチン様ホルモン / プロクラクン / ホルモン特異性
Research Abstract

1. Purification of human prolactin (hPRL), human growth hormone (hGH) and flounder growth hormone (fGH) expressed in E. coli.
(1) Expression vectors for hPRL, hGH and fGH were constructed using Taq promotor and each cDNA of hPRL, hGH and fGH, and were introduced into E. coli. The transformed E. coli expressed large amouamount of each protein, about 20% of the total E. coli proteins.
(2) The recombinant hormones were extracted from the E. coli cells, refolded with gultathione, and were purified to homogeneity by gel filtration and anion exchange chromatography.
2. Analysis of biological activities of recombinant PRL and GHs
(1) Biological activities of the recombinant hPRL and hGH were assayed for ability to stimulate the growth of Nb2 lymphoma cells and for ability to induce N-acetyllactosamine synthase in mammary grand cells, respectively. The biological activities of both recombinant hormones were equivalent to those of the corresponding native hormones.
(2) Biological activity of the recombinant fGH was assayed for growth promoting activity for rainbow trout. Although the fGH lacks the region composing 13-14 amino acid residues near C-terminal in other GHs including yellow tail GH, the recombinant fGH has exhibited equal activity to a recombinant yellow tail GH. This suggests that the region lacking in the fGH is not essential for the growth promoting activity.
3. Analysis of primary structure of prolactin-like hormones in bovine placenta. Four novel cDNA clones for prolactin-like proteins, bPLP-I, bPLP-II, bPLP-III and bPLP-IV were isolated from a bovine placental cDNA library, and their complete nucleotide sequences were determined. One of the clones, bPLP-IV, lacks the two C-terminal cysteine residues which are completely conserved in all the known members of GH/PRL gene family, suggesting that the two C-terminal cysteine residues may not be essential for lactogenic activity.

Report

(3 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report
  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] Minoru Tanaka: "Isolation of a novel prolactinーlike cDNA clone from bovine placenta:occurrence of new family members" Biochim.Biophys.Acta. 1008. 193-197 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Minoru Yamakawa: "Effect of gene dosage on the expression of human growth hormone cDNA in Escherichia coli" Biochim.Biophys.Acta. 1009. 156-160 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Minoru Yamakawa: "Expression of new members of the prolactin growth hormone gene family in bovine placenta" J.Biol.Chem.265. 8915-8920 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Minoru Yamakawa: "Nucleotide sequence of cDNA and primary structure for hard tail growth hormone" Biochim.Biophys.Acta. 1087. 247-249 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Minoru Yamakawa: "Homology in 5'ーand 3'ーnoncoding regions of hard tail growth hormone cDNA indicates close phylognic relationship in teleostei" Mie Med.J.40. 327-335 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Minoru Tanaka: "A novel cDNA clone encoding a prolactinーlike protein that lacks the two Cーterminal cysteine residues isolated from bovine placenta" Biochim.Biophys.Acta. (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Minoru Tanaka et al: "Isolation of a novel prolactin-like cDNA clone from bovine placenta : occurrence of new family members" Biochim, Biophys. Acta. 1008. 193-197 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Minoru Yamakawa et al: "Effect of gene dosage of the expression of human growth hormone cDNA in Escherichia coli" Biochim, Biophys. Acta. 1009. 156-160 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Minoru Yamakawa et al: "Expression of new family members of the prolactin growth hormone gene family in bovine placenta" J. Biol. Chem.265(15). 8915-8920 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Minoru Yamakawa et al: "Nucleotide sequence of cDNA and primary structure for hard tail growth hormone" Biochim. Blophys. Aata. 1087. 247-249 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Minoru Yamakawa et al: "Homology in 5'- and 3' -noncoding regions of hard tail growth hormone cDNA indicates close phylogenic relationship in teleostei" Mie Med. J.40, (3). 327-335 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Minoru Tanaka et al: "A novel cDNA clone encoding a prolactin-like protein that lacks the two C-terminal cysteine residues isolated from the bovine placenta" ^iochim, Biophys. Acta. (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Minoru Yamakawa: "Expression of new members of the prolactin growth hormore gene family in bovine placenta" J.Biol.Chem.265. 8915-8920 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Minoru Yamakawa: "Nucleotide sequence of cDNA and primary structure for hard tail growth hormone" Biochim.Biophys.Acta. 1087. 247-249 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Minoru Yamakawa: "Homology in 5'ーand 3'ーnoncoding regions of hard tail growth hormore cDNA indicates close phylogenetic relationship in teleostei" Mie Med.J.40. 327-335 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Minoru Tanaka: "A novel cDNA clone encoding a prolactinーlike protein that lacks the two Cーterminal cysteine residues isolated from bovine placenta" Biochim.Biophys.Acta. (1991)

    • Related Report
      1990 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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