Molecular characteristics and physiological roles of phosphatases in calcifying matrix vesicles.

钙化基质囊泡中磷酸酶的分子特征和生理作用。

基本信息

  • 批准号:
    63480412
  • 负责人:
  • 金额:
    $ 3.2万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for General Scientific Research (B)
  • 财政年份:
    1988
  • 资助国家:
    日本
  • 起止时间:
    1988 至 1989
  • 项目状态:
    已结题

项目摘要

The molecular process leading to apatite formation in matrix vesicle(MV) during the initial phase of calcification has never been understood so far on the basis of functional behavior of the intravesicular phosphatases. The project has been promoted with two types of MV participating each in different modes of calcification.(1) MV from enchondral ossification process: The MV from calf epiphyseal cartilage fractionated finally through Percoll density gradient centrifugation showed the highest alkaline phosphatase(ALP) specific activity among other organelles. The phosphotransferase activity of the ALP is interpreted as an essential function in calcification in or by MV. Apparent heterogeneity of MV suggested from the unfixed ratio of acid phosphatase(ACP)/ALP activity did not permit to ascribe the ACP as a unique vesicular enzyme despite possible localization of non-lyzosomal ACP in MV.(2) MV from intramembranous ossification process: The cultured cell of osteoblastic clone MC3T3-EI from mouse calvaria was examined in respect of the MV formation. In-between the ending of the monolayer growth where ALP level elevates in consequence of the retinoic acid receptor genes expression and the starting of the multi-layer growth together with Ca deposit, has been shown to be critical timing for the MV release. Thus, the criteria for sampling of calcifying MV have been established.(3) Automated micro stopped flow apparatus for rapid phosphatase assay: The apparatus constructed was powerful in dealing with very limited amounts of MV preparation in case of the fractionation or kinetic assay. Taking advantage of the above new apparatus, the MV from the osteoblastlike cultured cells is proved to be fruitful experimental system.
在钙化初始阶段,基质囊泡(MV)中导致磷灰石形成的分子过程迄今尚未基于囊泡内磷酸酶的功能行为被理解。两种MV以不同的钙化模式参与了项目的推进。(1)内软骨成骨过程的碱性磷酸酶(ALP)比活性:经Percoll密度梯度离心分离最终分离的小牛骨骺软骨的碱性磷酸酶(ALP)比活性在其他细胞器中最高。ALP的磷酸转移酶活性被解释为MV中或由MV引起的钙化的重要功能。从酸性磷酸酶(ACP)/ALP活性的不固定比率来看,MV具有明显的异质性,尽管可能在MV中定位了非溶酶体ACP,但不能将ACP归因于独特的囊泡酶。(2)膜内骨化过程中的MV:用小鼠颅骨成骨细胞MC3T3-EI培养细胞,观察其MV的形成情况。由于维甲酸受体基因表达导致ALP水平升高,在单层生长结束和多层生长开始以及Ca沉积之间,已被证明是MV释放的关键时间。从而建立了钙化MV的采样标准。(3)用于快速磷酸酶测定的自动微停流装置:在分馏或动力学测定中,该装置在处理非常少量的MV制剂方面功能强大。利用上述装置,从成骨样培养细胞中提取MV是一种卓有成效的实验系统。

项目成果

期刊论文数量(26)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
高橋浩二郎,谷口茂彦: "全自動型微量ストップドフロ-装置の試作と、その液体クロマトグラフィ-用リアルタイムアマライザ-としての応用。" 生化学. 60. 1390-1394 (1988)
Kojiro Takahashi、Shigehiko Taniguchi:“全自动微量停流装置的原型生产及其作为液相色谱实时氨化器的应用。” 60. 1390-1394 (1988)。
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    0
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Takahashi,K.,Yanagida,A.,Shimidzu,M.,Shindo,H.and Taniguchi,S.: "Intracellular levels of epidermal growth factor reaceptor and acid/alkaline phosphatases in proliferating mouse osteoblastic MC3T3 and human epidermoid carcinoma A431 cells."
Takahashi,K.、Yanagida,A.、Shimidzu,M.、Shindo,H. 和 Taniguchi,S.:“增殖的小鼠成骨细胞 MC3T3 和人表皮样癌 A431 细胞中表皮生长因子受体和酸/碱性磷酸酶的细胞内水平。
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TANIGUCHI Shigehiko其他文献

TANIGUCHI Shigehiko的其他文献

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{{ truncateString('TANIGUCHI Shigehiko', 18)}}的其他基金

Three-dimensional molecular status of bone morphogenesis as revealed from molecular biological analyzes.
分子生物学分析揭示骨形态发生的三维分子状态。
  • 批准号:
    02404072
  • 财政年份:
    1990
  • 资助金额:
    $ 3.2万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (A)
Development of an Automated Dual Type of Micro Stopped-Flow/Flow-Injection Apparatus Applicable for Detection of Minute Alteration of Inorganic Phosphate or Phosphoric Group in Protein Level
一种适用于检测蛋白质水平中无机磷酸盐或磷基微小变化的自动化双型微停流/流动注射装置的研制
  • 批准号:
    01870077
  • 财政年份:
    1989
  • 资助金额:
    $ 3.2万
  • 项目类别:
    Grant-in-Aid for Developmental Scientific Research (B).
Identification and early detection of oral cancer by the use of anti-EGF-receptor monoclonal antibody.
使用抗 EGF 受体单克隆抗体识别和早期检测口腔癌。
  • 批准号:
    61480388
  • 财政年份:
    1986
  • 资助金额:
    $ 3.2万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (B)

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