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Determination of different gonadotropin synthesis mechanisms in trophoblast and choriocarcinoma cells

Research Project

Project/Area Number 63570133
Research Category

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

Allocation TypeSingle-year Grants
Research Field Pathological medical chemistry
Research InstitutionFaculty of Pharmaceutical Sciences, Nagasaki University

Principal Investigator

SAKAKIBARA Ryuzo  Faculty of Pharm. Sci. Nagasaki Univ. Associate Prof., 薬学部, 助教授 (30127229)

Co-Investigator(Kenkyū-buntansha) ISHIGURO Masatsune  Faculty of Pharm. Sci. Nagasaki Univ. Prof., 薬学部, 教授 (90038274)
今村 定臣  長崎大学, 医学部附属病院, 講師 (50136644)
Project Period (FY) 1988 – 1989
Project Status Completed (Fiscal Year 1989)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1989: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1988: ¥1,500,000 (Direct Cost: ¥1,500,000)
KeywordsHuman chorionic gonadotropin / hCG / Biosynthesis of hCG / hCG subunits / Glycoprotein hormone / BeWo cell / Processing of sugar chain / Choriocarcinoma / ヒト胎盤絨毛性ゴナドトロビン / hCG生合成 / hCGサブユニットの解離会合 / BeWo細胞のhCG産生
Research Abstract

Human chorionic gonadotropin (hCG) is a glycoprotein hormone consisting of noncovalently bonded subunits, alpha and beta. Both subunits contain two Asn(N)-linked sugar chains, in addition, the beta-subunit contains four Ser(O)-linked sugar chains. It has been suggested that altered glycosylation is induced in both alpha- and beta-subunits of hCG produced by choriocarcinoma. The objective of this study was to determine the nature of the structural differences of hormones synthesized in the cells and secreted as well as the uncombined subunits in normal trophoblasts and a choriocarcinoma cell line (BeWo).
The results obtained from this study are as follows:
1. Intracellular forms of hCG subunits in normal trophoblast cells were analyzed by immunobinding method. It was revealed that hCG subunits accumulate in trophoblast cells as predominant intermediates (immature forms) containing high-mannose sugar chains. On the other hand, Ser-linked sugar chain was not found in the immature beta-subunit.
2. The immature hCG subunits were purified, separately, and their physicochemical properties were determined. N-terminal amino acid sequence of each immature subunit was identical to respective mature form. The purified immature subunits were recombined each other and reconstituted an immature hCG. The apparent molecular weights of the immature hCG found in trophoblast cells as well as hCG reconstituted with the purified immature subunits were larger than those expected. It was suggested that the immature hCG is present in trophoblast cells as an oligomer.
3. The structural characteristics of hCG and its subunits in choriocarcinoma, BeWo cells, were compared to those of normal trophoblast cells. It was found that both protein and sugar chain structures of hCG subunits of BeWo cells were differed from those of trophoblast cells.

Report

(3 results)
  • 1989 Annual Research Report   Final Research Report Summary
  • 1988 Annual Research Report
  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] 富永伸明: "Existence of Associated,Non-associted and Oligomeric Forms of Human Chorionic Gonadotropin Subunits in Placental Cells" J,Biochem.105. 992-997 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] 境晶子: "Human Chorionic Gonadotropin-Ricin A Chain Hybrid Protein:A Hormone Analog for the Study of Signal Transduction" J.Biochem.105. 275-280 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] 武内喜茂: "Synthesis and Secretion of Human Chorionic Gonadotropin and its Subunits in Choriocarcinoma Cells:A Comparative Study with Normal Placental Cells" Mol.Cell.Endocrinol.(1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] 榊原隆三: "Effective Purification of Human Chorionic Gonadotropin and its Subunits from Pregnant Women´s Urinary Peptides adsorbed on Reverse-Phase Resin" Chem.Pharm.Bull.(1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] 榊原隆三: "A Nicked β-Subunit of Human Chorionic Gonadotropin Purified from Pregnancy Urine" J,Biochem.(1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Nobuaki Tominaga, Ryuzo Sakakibara, Yoshihiro Yokoo and Masatsune Ishiguro: "Existence of Associated, Non-Associated, and Oligomeric Forms of Human Chorionic Gonadotropin Subunits in Placental Extracts" J. Biochem.105. 992-997 ((1989))

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Akiko Sakai, Ryuzo Sakakibara and Masatsune Ishiguro: "Human Chorionic Gonadotropin-Ricin A Chain Hybrid Protein; A Hormone Analog for the Study of Signal Transduction" J. Biochem. 105, 275-280 (1989).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Yoshishige Takeuchi, Ryuzo Sakakibara and Masatsune Ishiguro: "Synthesis and Secretion of Human Chorionic Gonadotropin and its Subunits in Choriocarcinoms Cells: A Comparative Study with Normal Placental Cells" Mol. Cell. Endocrinol. (1990).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Ryuzo Sakakibara, Syuichi Miyazaki, Toshiaki Shigemura, Nobuaki Tominaga, Akiko Sakai and Masatsune Ishiguro: "Effective Purification of Human Chorionic Gonadotropin and its Subunits from Pregnant Women's Urinary Peptides adsorbed on Reverse-Phase Resin" Chem. Pharm. Bull. (1990).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Ryuzo Sakakibara, Syuichi Miyazaki and Masatsune Ishiguro: "A nicked beta-subunit of Human Chorionic Gonadotropin Purified from Pregnancy Urine" J. Biochem. (1990).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] 武内喜茂: "Synthesis and Secretion of human Chorionic Gonadotropin and its Subunits in Choriocarcinoma cells:A comparative study with nornal placental cells" Mol.Cell.Endocrinol.(1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] 榊原隆三: "Effective Purification of Human Chorionic Gonadotropin and its subunits from Pregnant women's Urinary poptides adsorbed on Reverse-Phase Resin" Chem.Pharm.Bull.(1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] 榊原隆三: "A Nicked β-subunit of Human Chorionic Gonadotropin Purified from Pregnancy Urine" J.Biochem.(1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] 境晶子: J.Biochem. 105. 275-280 (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] 冨永伸明: J.Biochem. (1989)

    • Related Report
      1988 Annual Research Report

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Published: 1988-04-01   Modified: 2016-04-21  

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