Whole Statue of Recognition by Enzyme Toward Substarates with Vague Signals

酶识别模糊信号底物的完整雕像

• 批准号: 13680718
• 负责人: OGISHIMA Tadashi
• 金额: $ 2.24万
• 依托单位: Kyusyu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13680718/
• 关键词: processing; mitochondria; protease; substrate-recongnition; precursors; プロセシングペプチダーゼ; 基質認識機構; タンパク質分解; 光親和性標識

(1) Structural information was obtained using fluorescence resonance energy transfer measurement (FRET). A series of the peptide substrate with different intervening lengths between the distal and proximal argines were synthesized and then covalently attached with the fluorescence acceptor and donnaor. When the substrates were bound to the enzyme, essentially the same distances between the probes were obtained. Such common length was also obtained when when FRET was conducted between the substrates and enzyme.(2) Among many precursor proteins some proteins lack the proximal arginine residue. Such precursors were still cleaved by mitochondrial processing peptidase (MPP). When the distal residue was changed to arginine, the processing efficiency was slightly elevated. This indicates that some interactions are still present between the S2-P2 positions.(3) MPP was shown to cleave the internal peptide bond of a fusion protein. Although the fusion protein had established the individual structure, MPP could attacked the peptide pond in a loosely structured domain.(4) The interaction between MPP and the substrates around regions other than the cleavage site was investigated using FRET measurement. The results indicated that a portion downstream from the cleavage site about 10 amino acids could emerge from the MPP pocket.

（1）使用荧光共振能量传递测量（FRET）获得结构信息。合成了远端和近端精氨之间的一系列具有不同中间长度的肽底物，然后与荧光受体和Donnaor共同附着。当底物与酶结合时，获得了探针之间的基本相同距离。当在底物和酶之间进行FRET时，还可以获得这种常见的长度。（2）在许多前体蛋白中，有些蛋白质缺乏近端精氨酸残基。这种前体仍然通过线粒体加工肽酶（MPP）裂解。当将远端残基更改为精氨酸时，加工效率略有提高。这表明S2-P2位置之间仍然存在一些相互作用。（3）MPP被证明可以切割融合蛋白的内部肽键。尽管融合蛋白已经建立了个体结构，但MPP可以在一个松散结构化的结构域中攻击肽池。（4）使用FRET测量研究了MPP与裂解位点以外的区域周围其他区域之间的相互作用。结果表明，从MPP袋中出现了大约10个氨基酸的裂解部位下游的一部分。

项目成果

Kojima et al.: "Recognition of Mitochondrial Protein Precursor Lacking Arginine at Position 2 by Mitochondrial Processing Peptidase"J.Biochem.. 130. 497-502 (2001)

Kojima 等：“通过线粒体加工肽酶识别位置 2 缺乏精氨酸的线粒体蛋白前体”J.Biochem.. 130. 497-502 (2001)

Kojima, K., Kitada, S., Ogishima, T., Ito, A.: "A Proposed Common Structure of Substrates Bound to Mitochondrial Processing Peptidase"J.Biol.Chem.. 276. 2115-2121 (2001)

Kojima, K.、Kitada, S.、Ogishima, T.、Ito, A.：“与线粒体加工肽酶结合的底物的拟议共同结构”J.Biol.Chem.. 276. 2115-2121 (2001)

Kojima, K., Yamazaki, E., Kitada, S., Ogishima, T., Ito, A.: "Recognition of Mitochondrial Protein Precursor Lacking Arginine at Position -2 by Mitochondrial Processing Peptidase"J.Biochem.. 130. 497-502 (2001)

Kojima, K.、Yamazaki, E.、Kitada, S.、Ogishima, T.、Ito, A.：“通过线粒体加工肽酶识别位置 -2 处缺乏精氨酸的线粒体蛋白前体”J.Biochem.. 130. 497

Kojima., et al.: "Recongnition of Mitochondrial Protein Precursor Lacking Arginine at Position 2 by Mitochondrial Processing Peptidase"J. Biohem.. 130. 497-502 (2001)

Kojima., et al.：“通过线粒体加工肽酶识别位置 2 缺乏精氨酸的线粒体蛋白前体”J.

Kojima, K., Yamasaki, E., Kitada, S., Ogishima, T., Ito A: "Recognition of Mitochondrial Protein Precursor Lacking Arginine at Position -2 by Mitochondrial Processing Peptidase"J. Biochem.. 130. 497-502 (2001)

Kojima, K.、Yamasaki, E.、Kitada, S.、Ogishima, T.、Ito A：“线粒体加工肽酶对 -2 位缺乏精氨酸的线粒体蛋白前体的识别”J。