Discovery of chaperone-type nucleoside diphosphate kinase activity in Hsp70 and proteasome and its pathophysiological function in these proteins

Hsp70 和蛋白酶体中分子伴侣型核苷二磷酸激酶活性的发现及其在这些蛋白质中的病理生理功能

基本信息

项目摘要

Hsp70 is a multifunctional molecular chaperone whose interactions with protein substrates are regulated by ATP hydrolysis and ADP-ATP exchange. In the period granted by this foundation, we found that, in addition to ATPase activity, purified Hsp70 and 20S proteasome, a new family of N-terminal nucleophile hydrolases, free from nucleoside diphosphate (NDP) kinase, exhibit intrinsic ADP-ATP exchange activity. The rate constants for ATP hydrolysis and ATP synthesis of these proteins were in a similar range at the optimum pH of 7.5-8.5 in the presence of 5 mM ATP and 0.5 mM ADP.Both Hsp70 and 20S proteasome exhibited a considerably strict preference for ATP as a phosphate donor, and a biased substrate specificity, unlike NDP kinase. During the reaction, both proteins formed acid-labile autophosphorylated intermediates and nucleoside diphosphate-dependent dephosphorylation of the latters then occurred. These properties are not identical but similar to those of NDP kinase, and are not similar to those of adenylate kinase and ATP synthase. The 20S proteasome is composed of numerous low molecular mass subunits arranged in a stack of four rings, each containing seven different α- or β-subunits. Among these subunits, we identified that the C5 in the β-type and the C8 in the α-type subunits were autophosphorylated during the γ-phosphate transfer reaction and were photoaffinity labeled with 8-azido-[α-^<32>P] ATP, suggesting that the C5 and C8 subunits of the proteasome are responsible for the NDP kinase-like activity. We are now trying to identify the active sites of NDP kinase in these proteins and also try to identify the role of NDP kinase in the chaperone activity of Hsp70 and the conformational modification of substrates in the processing of proteolysis by 20S proteasome.
热休克蛋白70是一种多功能的分子伴侣,其与蛋白质底物的相互作用受ATP水解和ADP-ATP交换的调节。在该基金会的资助下,我们发现,除了ATP酶活性外,纯化的Hsp 70和20 S蛋白酶体,一个新的N-末端亲核水解酶家族,不含核苷二磷酸(NDP)激酶,表现出内在的ADP-ATP交换活性。在5 mM ATP和0.5 mM ADP存在下,这些蛋白质的ATP水解和ATP合成速率常数在最佳pH 7.5-8.5范围内相似。与NDP激酶不同,Hsp 70和20 S蛋白酶体都表现出对ATP作为磷酸供体的相当严格的偏好,并且具有偏向的底物特异性。在反应过程中,这两种蛋白质形成酸不稳定的自磷酸化中间体和核苷二磷酸依赖的去磷酸化的后者,然后发生。这些性质与NDP激酶的性质不相同但相似,并且与腺苷酸激酶和ATP合酶的性质不相似。20 S蛋白酶体由许多低分子量亚基组成,排列成四个环的堆叠,每个环包含七个不同的α-或β-亚基。在这些亚基中,我们鉴定了β-型亚基中的C5和α-型亚基中的C8在γ-磷酸转移反应中被自磷酸化,并被8-叠氮基-[α-^ P] ATP光亲和标记<32>,表明蛋白酶体的C5和C8亚基负责NDP激酶样活性。我们正在试图确定NDP激酶在这些蛋白中的活性位点,并试图确定NDP激酶在Hsp 70的伴侣活性中的作用以及20 S蛋白酶体蛋白水解过程中底物的构象修饰。

项目成果

期刊论文数量(47)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Hiroshi Mori, et al.: "14-3-3 τ associates with a translational control factor FKBP 12-rapamycin-associated protein in T cells after stimulation by pervanadate."FEBS Lett.. 467(1). 61-64 (2000)
Hiroshi Mori 等人:“在过钒酸盐刺激后,14-3-3 τ 与 T 细胞中的翻译控制因子 FKBP 12-雷帕霉素相关蛋白相关。”FEBS Lett.. 467(1) (2000)。 )
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Hiroshi Mori: "14-3-3 associates with a translational control ractor FKBP12-rapamycin-associated protein in T cells after stumulation by pervanadate"FEBS Lett.. 467(1). 61-64 (2000)
Hiroshi Mori:“在过钒酸盐刺激后,14-3-3 与 T 细胞中的翻译控制因子 FKBP12-雷帕霉素相关蛋白相关”FEBS Lett.. 467(1)。
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Hidehiro Takahashi et al.: "Increases levels of ε and γ isoforms of 14-3-3 proteins in cerebrospinal fluid in patients with Creutzfeldt-Jakob disease."Clin.& Diag.Lab.Immunol.. 6(6). 983-985 (1999)
Hidehiro Takahashi 等人:“增加克雅氏病患者脑脊液中 14-3-3 蛋白的 ε 和 γ 同工型水平。”Clin.& Diag.Lab.Immunol.. 6(6)。 985 (1999)
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木戸博: "分子シャペロンによるタンパク質の立体構造の管理とタンパク質分解"石浦章一 編(シュプリンガーフェアラーク). 12 (2000)
Hiroshi Kido:“分子伴侣对蛋白质 3D 结构和蛋白水解的管理”,Shoichi Ishiura 编辑(Springer Verlag)12(2000)。
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矢野仁康: "分子シャペロンと蛋白質分解酵素に認められた新機能、ヌクレオシド2リン酸キナーゼ型酵素活性"生化学. 72(1). 41-45 (2000)
Hiroyasu Yano:“分子伴侣和蛋白水解酶中发现的新功能:核苷二磷酸激酶型酶活性”生物化学 72(1) (2000)。
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KIDO Hiroshi其他文献

KIDO Hiroshi的其他文献

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{{ truncateString('KIDO Hiroshi', 18)}}的其他基金

Allergy research prospect with new paradigm open up by the new sensitive allergen microarray
新型敏感过敏原微阵列开辟过敏研究新范式
  • 批准号:
    17K19662
  • 财政年份:
    2017
  • 资助金额:
    $ 9.34万
  • 项目类别:
    Grant-in-Aid for Challenging Research (Exploratory)
Study on the pathogenesis of severe influenza virus infection associated with cytokine storm and its effective treatment options
细胞因子风暴相关重症流感病毒感染发病机制及有效治疗方案研究
  • 批准号:
    16H05348
  • 财政年份:
    2016
  • 资助金额:
    $ 9.34万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Discovery of antigen-specific low affinity IgE in cord blood and the mechanisms of affinity maturation after birth for prevention of allergy
脐带血中抗原特异性低亲和力 IgE 的发现及其出生后亲和力成熟预防过敏的机制
  • 批准号:
    15K15371
  • 财政年份:
    2015
  • 资助金额:
    $ 9.34万
  • 项目类别:
    Grant-in-Aid for Challenging Exploratory Research
Screening of Flu Alarmin biomarkers of severe influenza virus infection and their confirmation in animal model
重症流感病毒感染Flu Alarmin生物标志物的筛选及动物模型验证
  • 批准号:
    25670466
  • 财政年份:
    2013
  • 资助金额:
    $ 9.34万
  • 项目类别:
    Grant-in-Aid for Challenging Exploratory Research
Pathogenicity of multiple organ failure induced by seasonal and highly pathogenic avian influenza virus infection and its therapeutic options
季节性高致病性禽流感病毒感染致多器官功能衰竭的致病性及治疗选择
  • 批准号:
    24249059
  • 财政年份:
    2012
  • 资助金额:
    $ 9.34万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
Studies on real-time biomarker of illness severity inthe patients of critical illness and development of the diagnosis machine
危重症患者病情严重程度实时生物标志物研究及诊断机研制
  • 批准号:
    23659846
  • 财政年份:
    2011
  • 资助金额:
    $ 9.34万
  • 项目类别:
    Grant-in-Aid for Challenging Exploratory Research
Development of new therapeutics for a highly pathogenic influenza virus infection by inhibition of virus entry and multiplication
通过抑制病毒进入和增殖来开发高致病性流感病毒感染的新疗法
  • 批准号:
    21249061
  • 财政年份:
    2009
  • 资助金额:
    $ 9.34万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
Systematic extraction of vocabulary used to express auditory impressions of utterances
系统地提取用于表达话语听觉印象的词汇
  • 批准号:
    19500177
  • 财政年份:
    2007
  • 资助金额:
    $ 9.34万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Development of voice montage system.
开发语音蒙太奇系统。
  • 批准号:
    16300061
  • 财政年份:
    2004
  • 资助金额:
    $ 9.34万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Development of new influenza virus treatments based an the findings of triggering proteases for influenza virus entry into the cells and on the findings of protease-activating receptors
基于触发流感病毒进入细胞的蛋白酶的发现以及蛋白酶激活受体的发现,开发新的流感病毒治疗方法
  • 批准号:
    13557014
  • 财政年份:
    2001
  • 资助金额:
    $ 9.34万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)

相似国自然基金

抑素蛋白(prohibitin)1调控蛋白酶激活受体(protease-activated receptor)1内化转运及降解的功能和机制
  • 批准号:
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三角帆蚌丝氨酸蛋白酶(serine protease)基因的克隆、表达调控与功能研究
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CAREER: Next-generation protease inhibitor discovery with chemically diversified antibodies
职业:利用化学多样化的抗体发现下一代蛋白酶抑制剂
  • 批准号:
    2339201
  • 财政年份:
    2024
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    $ 9.34万
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    Continuing Grant
The underlying dynamic exchange that dictates serine protease function
决定丝氨酸蛋白酶功能的潜在动态交换
  • 批准号:
    2332239
  • 财政年份:
    2024
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    $ 9.34万
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Genetic studies of a pleiotropic transmembrane protease: insight from color variation in non-model organisms
多效性跨膜蛋白酶的遗传研究:从非模型生物体颜色变化中获得洞察
  • 批准号:
    10754001
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    2023
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    $ 9.34万
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Protease Resistant Growth Factor Nanoparticles for Chronic Wound Healing
用于慢性伤口愈合的蛋白酶抗性生长因子纳米颗粒
  • 批准号:
    10593195
  • 财政年份:
    2023
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    $ 9.34万
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The Serine Protease HTRA1 Antigen: A Gateway to Elucidating Membranous Nephropathy Pathogenesis and the Targeting of Antigen Epitopes
丝氨酸蛋白酶 HTRA1 抗原:阐明膜性肾病发病机制和抗原表位靶向的途径
  • 批准号:
    10740614
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    2023
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    $ 9.34万
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Establishment of efficient virus isolation by protease expression cells
通过蛋白酶表达细胞建立有效的病毒分离
  • 批准号:
    22KJ0023
  • 财政年份:
    2023
  • 资助金额:
    $ 9.34万
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    Grant-in-Aid for JSPS Fellows
Candida albicans Sap6 dysregulates host epithelial protease-antiprotease expression
白色念珠菌 Sap6 失调宿主上皮蛋白酶-抗蛋白酶表达
  • 批准号:
    10739848
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    2023
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Protease-activated-receptor-2 antagonists for treatment of migraine pain
蛋白酶激活受体 2 拮抗剂治疗偏头痛
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    10602826
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Development of innovative drug discovery based on SARS-CoV-2 3CL protease inhibitor YH-53
基于SARS-CoV-2 3CL蛋白酶抑制剂YH-53的创新药物研发
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    23H02614
  • 财政年份:
    2023
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    $ 9.34万
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    Grant-in-Aid for Scientific Research (B)
Deciphering the role of protease-activated receptor 2 in immuno-oncology
破译蛋白酶激活受体 2 在免疫肿瘤学中的作用
  • 批准号:
    488385
  • 财政年份:
    2023
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    $ 9.34万
  • 项目类别:
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