Molecular mechanism of processing of precursor proteins in organelles derived from parasitic organisms

寄生生物细胞器中前体蛋白加工的分子机制

• 批准号: 10480171
• 负责人: ITO Akio
• 金额: $ 8万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10480171/
• 关键词: processing peptidase; processing; precursor protein; mitochondria; signal recognition; parasite; X-ray analysis; peptide substrate; X線構造解析; 前駆体タンパク質; ペプチド基質; ヒドロゲノソーム

Mechanism of processing of precursor proteins in organelles derived from parasitic organisms.(1) Mitochondrial processing peptidase : Systematic and quantitative analyses, using various synthetic peptides based on the amino acid sequence of the extension peptides of the precursors, demonstrated that a combination of residues around the cleavage site, including the proximal arginine and residues at positions 1, 2, and 3, and the distal basic amino acids appears to be responsible for recognition of the substrates and for determination of the cleavage site by MPP.Functional amino acid residues in β-MPP were determined by site-directed mutation experiments. Shortening of the glycine-rich stretch in α-MPP led to a dramatic reduction of interaction between the enzyme and substrate peptides and cleavage reaction. Fluorescence resonance energy transfer (FRET) measurements indicate that the N-terminal portion and the portion around the cleavage site of the extension peptides interact with specific sites in the MPP molecule. The structures of recombinant yeast MPP and a cleavage-deficient mutant of MPP complexed with synthetic signal peptides were determined using X-ray crystallographic techniques.(2) MPP-like proteases of protozoan : cDNAs of MPP-like proteases of trypanosoma was obtained and the amino acid sequence was deduced.

寄生生物细胞器中前体蛋白的加工机制。(1)线粒体加工肽酶：使用基于前体的延伸肽的氨基酸序列的各种合成肽的系统和定量分析证明，切割位点周围的残基的组合，包括近端精氨酸和位置1、2和3的残基，β-MPP的功能性氨基酸残基由酶切位点确定，酶切位点由酶切位点确定，酶切位点由酶切位点确定。定向突变实验α-MPP中富含甘氨酸的片段的缩短导致酶与底物肽之间的相互作用和切割反应的显著减少。荧光共振能量转移（FRET）的测量表明，N-末端部分和周围的延伸肽的切割位点的部分与MPP分子中的特定位点相互作用。利用X射线晶体学技术测定了重组酵母MPP和与合成信号肽复合的MPP切割缺陷突变体的结构。(2)原虫类MPP蛋白酶：获得了锥虫类MPP蛋白酶的cDNA，并推导了其氨基酸序列。

项目成果

小島克彦 等: "Cooperative Formation of a Substrate-Binding Pocket by a- and b- subunits of Mitochondrial Processing Peptidase"J.Biol.Chem.. 273. 32542-32546 (1998)

Katsuhiko Kojima 等人：“线粒体加工肽酶的 a- 和 b- 亚基协同形成底物结合袋”J.Biol.Chem.. 273. 32542-32546 (1998)

小島克彦 等: "A Proposed Common Structure of Substrates Bound to Mitochondrial Processing Peptidase"J.Biol.Chem.. 276. 2115-2121 (2001)

Katsuhiko Kojima 等人：“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)

伊藤明夫: "タンパク質の細胞内輸送とプロセシング"化学工業. 51. 51-57 (2000)

Akio Ito：“蛋白质的细胞内运输和加工” Kagaku Kogyo。51. 51-57 (2000)

伊藤明夫: "タンパク質分解-分子機構と細胞機能"東京化学同人. 236 (2000)

伊藤昭夫：“蛋白水解-分子机制和细胞功能”东京化学同人236（2000）。

小島克彦 等: "Cooperative Formation of a Substrate-Binding Pocket by α- and β-subunits of Mitochondrial Processing Peptidase." J.Biol.Chem.273・49. 32542-32546 (1998)

Katsuhiko Kojima 等人：“线粒体加工肽酶的 α 和 β 亚基协同形成底物结合袋。”J.Biol.Chem.273·49（1998）。