Intracellular Processing, Activation, and Sorting of Lysosomal Cathepsins

溶酶体组织蛋白酶的细胞内加工、激活和分选

基本信息

  • 批准号:
    01580195
  • 负责人:
  • 金额:
    $ 1.28万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)
  • 财政年份:
    1989
  • 资助国家:
    日本
  • 起止时间:
    1989 至 1990
  • 项目状态:
    已结题

项目摘要

Lysosomal cathepsins are considered to play nd important role in intracellular protein degradation. Like most lysosomal acid hydrolases, cathepsins are synthesized as the glycosylated preproenzymens on membrane-bound ribosomes of endoplasmic reticulum and the higher-molecular-weight precursors are subsequently converted into the mature enzymes posttranslationally by limited proteolysis.In my studies, I have found the following evidence : (1) The latent proforms of cathepsins B, H, L, and D were identified by immunoblot analysis in the rat liver microsomal lumen, and the in vitro activation experiments with liver microsomal contents revealed that latent procathepsins are converted to the active forms of mature enzymes under acidic conditions. (2) The proteolytic processing events, and the activation of the enzymes were strongly inhibited by pepstatin, therefore, suggesting that cathepsin D, a major lysosomal aspartic proteinase is involved in the propeptide-processing of procathepsins. … More (3) To further characterize these intracellular processing and activation mechanisms for the lysosomal cathepsins, microsomal procathepsin B was highly purified by the Con A-Sepharose, Sepharose-Gly-Phe-GlySc, and the anti-cathepsin B IgG Sepharose and then this proenzyme was incubated with rat liver tritosomal contents in the presence of several proteinase inhibitors. The result confirmed that the lysosomal cathepsin D is indeed a processing proteinase of procathepsin B. Furthermore, the processed form of procathepsin after the digestion with cathepsin D was isolated and subjected to the determination of the NH_2-terminal sequence of the molecule. The determined NH_2-terminal sequence of procathepsin B revealed to be Pro-66 which is 14 amino acids upstream of the NH_2-terminus of the mature cathepsin B, suggesting that the procathepsin B would undergo multiple processing step during intracellular transport. (4) The multicopy plasmid containing the cathepsin L cDNA has been introduced into the yeast to determine the intracellular sorting machinery of lysosomal cathepsin L. These in vivo and in vitro studies will help me to clarify the extracellular processing and activation mechanisms lysosomal cathepsins. Less
溶酶体组织蛋白酶被认为在细胞内蛋白质降解中起重要作用。与大多数溶酶体酸性水解酶一样,组织蛋白酶是在内质网膜结合核糖体上合成的糖基化前体,高分子量的前体随后通过有限的蛋白水解后转化为成熟的酶。(1)通过免疫印迹分析在大鼠肝微粒体管腔中鉴定组织蛋白酶B、H、L和D的潜伏前体,并且用肝微粒体内容物进行的体外活化实验揭示,在酸性条件下,潜伏的组织蛋白酶原被转化为成熟酶的活性形式。(2)胃蛋白酶抑制剂强烈抑制蛋白水解加工事件和酶的活化,因此,表明组织蛋白酶D,一种主要的溶酶体天冬氨酸蛋白酶参与前肽加工的前体蛋白酶。 ...更多信息 (3)为了进一步表征溶酶体组织蛋白酶的这些细胞内加工和活化机制,通过Con A-Sepharose、Sepharose-Gly-Phe-GlySc和抗组织蛋白酶B IgG Sepharose高度纯化微粒体组织蛋白酶原B,然后在存在几种蛋白酶抑制剂的情况下将该酶原与大鼠肝tritosomal内容物孵育。结果证实溶酶体组织蛋白酶D确实是组织蛋白酶原B的加工蛋白酶。此外,还分离了经组织蛋白酶D消化后的蛋白酶原,并测定了其氨基端序列。组织蛋白酶原B NH_2-端序列测定结果表明,组织蛋白酶原B NH_2-端上游14个氨基酸为Pro-66,提示组织蛋白酶原B在胞内转运过程中可能经历多个加工步骤。(4)将含有组织蛋白酶L cDNA的多拷贝质粒导入酵母,以确定溶酶体组织蛋白酶L的细胞内分选机制。这些在体内和体外的研究将帮助我阐明细胞外加工和激活机制溶酶体组织蛋白酶。少

项目成果

期刊论文数量(20)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Nishimura,Y.: "Inhibition of intracellular sorting and processing of lysosomal cathepsins H ahd L at reduced temperature in primary cultures of rat hepato cytes." Archives of Biochemistry and Biophysics. 283. 458-463 (1990)
Nishimura,Y.:“在大鼠肝细胞原代培养物中,在低温下抑制溶酶体组织蛋白酶 H 和 L 的细胞内分选和加工。”
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Nishimura,Y: "Intracellulan processing and activation of lysosomal cathepsins" Acta Histochemica et Cytochamica. 23. 53-64 (1990)
Nishimura,Y:“溶酶体组织蛋白酶的细胞内加工和激活”《组织化学与细胞化学学报》。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Oda,K.: "Brefeldin A inhibits the targeting of cathepsin D and cathepsin H to lysosomes in rat hepatocytes" Biochem.Biophys.Res.Commun.163. 220-225 (1989)
Oda,K.:“布雷菲德菌素 A 抑制组织蛋白酶 D 和组织蛋白酶 H 靶向大鼠肝细胞中的溶酶体”Biochem.Biophys.Res.Commun.163。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Nishimura,Y: "Intracellular processing and activation of lysosomal cathepsin L.in“Intracellular proteolysis,Mechanisms,and Regulations"(eds.Katunuma,N.,Kominami,E.)" SpringerーVerlag/Japan Sci.Soc., 13 (1989)
Nishimura, Y:“细胞内蛋白水解、机制和调控中溶酶体组织蛋白酶 L. 的细胞内加工和激活”(Katunuma, N.、Kominami, E. 编)”Springer-Verlag/Japan Sci.,13( 1989)
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

NISHIMURA Yukiko其他文献

NISHIMURA Yukiko的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('NISHIMURA Yukiko', 18)}}的其他基金

A Sociolinguistic Study of Expanding Digital Communication in Japanese: In View of Ageing
扩大日语数字交流的社会语言学研究:考虑到老龄化
  • 批准号:
    24520479
  • 财政年份:
    2012
  • 资助金额:
    $ 1.28万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
A Sociolinguistic Japan-UK Joint Research of the Japanese Language Online and its Speakers' Interactional Linguistic Behaviour
日英社会语言学在线日语及其使用者互动语言行为的联合研究
  • 批准号:
    21520448
  • 财政年份:
    2009
  • 资助金额:
    $ 1.28万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Study for Human resource for Orphan Drug Development and Rare Disease (NANBYO) Research in Japan
日本孤儿药开发和罕见病(NANBYO)研究人力资源研究
  • 批准号:
    21730288
  • 财政年份:
    2009
  • 资助金额:
    $ 1.28万
  • 项目类别:
    Grant-in-Aid for Young Scientists (B)
Study for the possible creation of new orphan drug market based on the start-ups from the universities in Japan
基于日本大学初创企业的新孤儿药市场创建可能性研究
  • 批准号:
    18730241
  • 财政年份:
    2006
  • 资助金额:
    $ 1.28万
  • 项目类别:
    Grant-in-Aid for Young Scientists (B)

相似海外基金

DRAM2: on the crossroad between trans-Golgi network and lysosomes?
DRAM2:位于跨高尔基体网络和溶酶体之间的十字路口?
  • 批准号:
    2882796
  • 财政年份:
    2023
  • 资助金额:
    $ 1.28万
  • 项目类别:
    Studentship
Mechanotransduction from lysosomes drives sterile inflammation
溶酶体的力传导驱动无菌炎症
  • 批准号:
    479441
  • 财政年份:
    2023
  • 资助金额:
    $ 1.28万
  • 项目类别:
    Operating Grants
Role of microglial lysosomes in amyloid-A-beta degradation
小胶质细胞溶酶体在淀粉样蛋白-A-β降解中的作用
  • 批准号:
    10734289
  • 财政年份:
    2023
  • 资助金额:
    $ 1.28万
  • 项目类别:
Membrane trafficking to lysosomes
膜转运至溶酶体
  • 批准号:
    10620966
  • 财政年份:
    2023
  • 资助金额:
    $ 1.28万
  • 项目类别:
Lysosomes as the starting point of innate immune responses - importance of lysosomal environment -
溶酶体作为先天免疫反应的起点 - 溶酶体环境的重要性 -
  • 批准号:
    23H00366
  • 财政年份:
    2023
  • 资助金额:
    $ 1.28万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
The role of chloride in supporting the degradative capacity of phagosomes and lysosomes
氯化物在支持吞噬体和溶酶体降解能力中的作用
  • 批准号:
    RGPIN-2022-04485
  • 财政年份:
    2022
  • 资助金额:
    $ 1.28万
  • 项目类别:
    Discovery Grants Program - Individual
Microglial lysosomes and selective neuronal vulnerability
小胶质细胞溶酶体和选择性神经元脆弱性
  • 批准号:
    10829767
  • 财政年份:
    2022
  • 资助金额:
    $ 1.28万
  • 项目类别:
The role of chloride in supporting the degradative capacity of phagosomes and lysosomes
氯化物在支持吞噬体和溶酶体降解能力中的作用
  • 批准号:
    DGECR-2022-00214
  • 财政年份:
    2022
  • 资助金额:
    $ 1.28万
  • 项目类别:
    Discovery Launch Supplement
Investigating mechanisms activating the selective autophagy of lysosomes
研究激活溶酶体选择性自噬的机制
  • 批准号:
    10386081
  • 财政年份:
    2022
  • 资助金额:
    $ 1.28万
  • 项目类别:
Investigating mechanisms activating the selective autophagy of lysosomes
研究激活溶酶体选择性自噬的机制
  • 批准号:
    10634497
  • 财政年份:
    2022
  • 资助金额:
    $ 1.28万
  • 项目类别:
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了