Molecular mechanism of perexisomal protein import into peroxisomes.

过氧化物酶体蛋白导入过氧化物酶体的分子机制。

• 批准号: 08680661
• 负责人: MIURA Satoshi
• 金额: $ 1.6万
• 依托单位: YOKOHAMA CITY UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08680661/
• 关键词: Peroxisome; Acyl-CoH oxidase; Uricase; Molecular chaperon; アシルCoAオキシダーゼ; アシル-CoAオキシダーゼ

To investigate the molecular mechanism of protein localization to peroxisome, we have found out several new facts as follows using in vitro import system of several peroxisomal proteins.1.Newly imported uricase in the in vitro import system quickly assembled to core structure found in the rat liver peroxisomes.2.We investigated the importance of proline residue near the C-terminal PTS-1 signal, which is frequently obserbed in the flanking regions of the protein-protein interaction sites.3.ATP hydrolysis is essential for acyl-CoA oxidase and uricase import into peroxisomes, and other three nucleotide tri-phosphates, such as GTP,CTP or UTP can substitute for ATP4.The molecular chaperone such as mitochondrial import stimulation factor (MSF) and HSC70, which are important for the mitochondrial protein translocation to mitochondria, has no essential role for the peroxisomal protein import into peroxisomes. We showed the presence of unidentified proteineous factor (S) essential for peroxisomal import.5.We developed the in vitro translocation of serine : pyruvate aminotransferase (SPT) into peroxisomes, and found that the association of peroxisomal SPT with the peroxisomal membrane cause a marked confomational change in the structure of this protein.

为了探讨蛋白质定位于过氧化物酶体的分子机制，我们利用几种过氧化物酶体蛋白的体外导入系统，发现了以下新的事实：1.新导入的尿酸酶在体外导入系统中快速组装成大鼠肝脏过氧化物酶体中发现的核心结构; 2.我们研究了PTS-1信号C端附近脯氨酸残基的重要性，其通常位于蛋白质-蛋白质相互作用位点的侧翼区域。3. ATP水解对于酰基-CoA氧化酶和尿酸酶输入过氧化物酶体是必需的，而其它三种核苷酸三磷酸，如GTP，CTP或UTP可替代ATP 4。线粒体输入刺激因子（MSF）和HSC 70等分子伴侣在线粒体蛋白向线粒体转运中起重要作用，对过氧化物酶体蛋白输入过氧化物酶体没有重要作用。5.我们建立了丝氨酸：丙酮酸氨基转移酶（serine：pyruvate aminotransferase，SPT）在体外转运到过氧化物酶体中的方法，发现SPT与过氧化物酶体膜的结合引起了该蛋白质结构的显著变化。

项目成果

Toshiaki Oda: "In vitro association with peroxisomes and confomational change of peroxisonal serineipyruvale/alanmeiglyoxylale aminolransferase in rat and haran livers" Biochem.Biophys.Res.Commun.228. 341-346 (1996)

Toshiaki Oda：“大鼠和哈兰肝脏中过氧化物酶体和过氧化物酶体丝氨酸丙酮酸/丙二醛氨基转移酶的构象变化的体外关联”Biochem.Biophys.Res.Commun.228。

T.Oda, T.Funai and S.Miura: "In vitro association with peroxisomes and conformational change of peroxisomal serine : pyruvate/alanine : glyoxylate aminotransferase in rat and human livers" Biochem.Biophys.Res.Commun.228. 341-346 (1996)

T.Oda、T.Funai 和 S.Miura：“大鼠和人类肝脏中过氧化物酶体和过氧化物酶体丝氨酸构象变化的体外关联：丙酮酸/丙氨酸：乙醛酸转氨酶”Biochem.Biophys.Res.Commun.228。

Toshiaki Oda, Tuneyoshi Funai and Satoshi Miura: "In Vitro Association with Peroxisomes and Confomational change of Peroxisomal Serine: Pyvuvate/Alanimc 1 Glyoxylate Aminotransferase in Rat and Human Livers" Biochemical and Biophysical Research Communicat

Toshiaki Oda、Tuneyoshi Funai 和 Satoshi Miura：“与过氧化物酶体的体外关联和过氧化物酶体丝氨酸的构象变化：大鼠和人类肝脏中的丙酮酸/Alanimc 1 乙醛酸转氨酶”生物化学和生物物理研究通讯