An analysis of mitochondrial transport mechanisms and physiological functions of mammalian molecular chaperone HSP60

哺乳动物分子伴侣HSP60线粒体转运机制及生理功能分析

• 批准号: 12670105
• 负责人: ITOH Hideaki
• 金额: $ 2.18万
• 依托单位: AkitaUniversity
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670105/
• 关键词: Molecular chaperone; Heat shock protein; Stress protein; Mitochondria; シグナルペプチド; ミトコンドリア移行

Prokaryotic chaperonin has been well characterized, and the crystal structure of the asymmetric GroEL-GroES-(ADP)_7 chaperonin complex has been shown by Xu et al. On the contrary, biochemical properties of mammalian chaperonin homolog (HSP60/10) have not yet fully understood.We have analyzed binding protein to the signal sequence of HSP60 using signal sequence affinity column chromatography and identified only one protein with molecular mass of 70kDa. The protein was identified as HSP70 on immunoblotting using an antibody against HSP70.We have also investigated import of HSP60 into mitochondria in vivo. The mRNA of HSP60 was.increased in the papilla of water- restricted rat kidneys for 3 or 5 days. On immunoblotting analysis, HSP60 was detected both in cytoplasm and mitochondria of normal rat kidney papilla. Although the quantity of the protein was decreased in the cytoplasm of water-restricted rat kidney papilla, the protein was increased in the mitochondria of the papilla.These results suggested that mammalian molecular chaperon HSP60 is located both in the cytoplasm and mitochondria under normal conditions, and that cytoplasmic HSP60 is imported into mitochondria under severe conditions through cytoplasmic HSP70.

原核生物伴侣蛋白已经得到了很好的表征，不对称GroEL-Groes-(ADP)_7伴侣蛋白复合体的晶体结构已经被Xu等人证明。相反，哺乳动物伴侣蛋白同源物(HSP60/10)的生化性质尚未完全了解。我们利用信号序列亲和柱层析分析了与HSP60信号序列结合的蛋白，仅鉴定出一个相对分子质量为70 kDa的蛋白。用抗HSP70的抗体进行免疫印迹分析，确定该蛋白为HSP70。我们还研究了体内HSP60进入线粒体的情况。限水3天或5天的大鼠肾脏乳头中HSP60的mRNA表达增加。免疫印迹分析显示，HSP60在正常大鼠肾乳头细胞胞浆和线粒体中均有表达。结果表明，正常情况下哺乳动物分子伴侣HSP60存在于细胞质和线粒体中，而在严酷条件下，细胞质HSP60通过细胞质HSP70进入线粒体。

项目成果

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Takahashi S.: "Functional interaction of the immunosuppressant mizoribine with the 14-3-3 protein"Biochem. Biophys. Res. Commun. 274. 87-92 (2000)

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Motoyama, S: "Hydrdrogen peroxide induces midzonal heat shock protein 72 and apoptosis in sinusoidal endothelial cells of hypoxic rat liver"Crit. Care Med.. 28. 1509-1514 (2000)

Motoyama, S：“过氧化氢诱导缺氧大鼠肝脏中区热休克蛋白 72 和肝窦内皮细胞凋亡”Crit。

Iwabuchi A.: "Specific preinduction of 60-kDa heat shcok protein (chaperonin homolog) by TRH does not protect clonic mucosa against acetic acid-induced lesion in rat"Digest. Dis. Sci. 45. 1480-1489 (2000)

Iwabuchi A.：“TRH 对 60-kDa 热休克蛋白（伴侣蛋白同系物）的特异性预诱导不会保护大鼠中阵挛粘膜免受乙酸诱导的病变”文摘。

Iwabuchi, A.: "Specific preinduction of 60-kDa heat shock protein (chaperonin homolog) by TRH does not protect clonic mucosa against acetic acid-induced lesion in rat"Digest. Dis. Sci.. 45. 1480-1489 (2000)

Iwabuchi, A.：“TRH 对 60-kDa 热休克蛋白（伴侣蛋白同源物）的特异性预诱导不会保护克隆粘膜免受乙酸诱导的大鼠病变”文摘。