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Studies on human osteopontin : isolation and characterization of osteopontin in milk and serum.

Research Project

Project/Area Number 09557062
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Pediatrics
Research InstitutionHOKKAIDO UNIVERSITY

Principal Investigator

KOBAYASHI Kunihiko  Hokkaido Univ. Sch. Med., Pro., 医学部, 教授 (60091451)

Co-Investigator(Kenkyū-buntansha) MAFUNE Naoki  Hokkaido Univ. Sch. Med., Assist., 医学部, 助手 (70241304)
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥9,200,000 (Direct Cost: ¥9,200,000)
Fiscal Year 1998: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1997: ¥5,900,000 (Direct Cost: ¥5,900,000)
Keywordsosteopontin / Ca-binding / human milk / human serum / molecular fragility / Ca結合蛋白 / 母乳 / 血清
Research Abstract

Osteopontin (hereafter called as OPN), a CaィイD12+ィエD1 binding protein, has been studied as a secreted highly acidic phophoglycoprotein with diverse functions in various tissue and species. It was firstly isolated from mineralized bone matrix and studied as a protein remodeling mineralized tissue. Since OPN is present in human milk, we considered that the OPN plays an important role in the osteogenesis in the developing infants. The present studies aimed to isolate and characterize OPN in human milk, serum and blood cells.
We have isolated osteopontin from human milk, prepared antibodies specific for it, and tried to establish a sandwich type EIA system determining osteopontin in serum. It was found that serum osteopontin but not milk osteopontin showed paradoxical phenomena in reaction with the antibodies, making no detectable precipitation in immunoelectrophoresis and no positive reaction in EIA, but showing positive reaction in Western blot and in an inhibition tests. Taking these phenomena and other circumstantial evidences into account, it is concluded that the serum osteopontin has a high molecular fragility, fragmenting into small peptides with few epitopes during incubation time. This fragility, though much lower extent, was also seen in milk osteopontin. Although the exact casual meaning of this fragility was not defined yet, it is conceivable that the fragility of OPN itself has some meaning in its physiological functions. Such possibility is being investigated in our laboratory.

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (9 results)

All Other

All Publications (9 results)

  • [Publications] 真船直樹: "血清オステオポンチン研究のピットホールー血清オステオポンチンの免疫生科学的不安定性"臨床病理. 45(補冊). 272 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 真船直樹: "臨床の求める測定値の精度ーククオグロブリン血症例 と血清オステオポンチン測定を例として"第74回北海道臨床衛生検査学会講演集. 26 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 真船直樹: "免疫電気泳動から推察するオステオポンチン測定のピットホール"生物物理化学. 42. 16 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Naoki Mafune, et al.: "A pitfall in study of osteopontin an immunochemical analysis of serum osteopontin"Jpn. J. Clin. Pathol.. 45. 272 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Naoki Mafune, et al.: "Acuracy of laboratory data necessary for clinical application - cryoglobulinameia and osteopontin"74th Hokkaido Rinsyoeiseikensa (Abst). 26 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Naoki Mafune, et al.: "A pitfall in the determination of osteopontin"Jpn. J. Electrophoresis. 42. 16 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 真船直樹: "血清オステオポンチン研究のピットホール-血清オステオポンチンの免疫生化学的不安定性" 臨床病理. 45(補冊). 272 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] 真船直樹: "臨床の求める測定値の精度-ククオグロブリン血症例と血清オステオポンチン測定を例として" 第74回北海道臨床衛生検査学会講演集. 26 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 真船直樹: "免疫電気泳動から推察するオステオポンチン測定のピットホール" 生物物理化学. 42. 16 (1998)

    • Related Report
      1998 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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