Structure and Function of Complex I of Mitochondrial Respiratory Chain

线粒体呼吸链复合物I的结构与功能

• 批准号: 01044089
• 负责人: MATSUBARA Hiroshi
• 金额: $ 5.63万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01044089/
• 关键词: Mitochondrial respiratory chain; Complex I; NADH dehydrogenase; Iron-sulfur protein; Chloroplast DNA; ミトコンドリア; 電子伝達系; 複合体I; 鉄イオウ蛋白質; 一次構造

Mitochondrial NADH : ubiquinone oxidoreductase (complex I) is the most complicated system in the respiratory chain. It consists of many subunits, some of which hold iron-sulfur clusters, but structural information is still limited. Two hydrophilic subfraction of bovine heart complex I were systematically resolved into individual polypeptides by chromatography. Three polypeptides (51, 24, and 9kDa) were isolated from the flavoprotein fraction (FP) of complex I, and the complete amino acid sequence of the 9kDa polypeptide was determined. The 9kDa polypeptide was composed of 75 amino acids with a molecular weight of 8437. This protein exhibited no obvious sequence similarity to other proteins. The iron-sulfur protein fraction (IP) of complex I was separated into eight polypeptides, 75, 49, 30, 20, 18, 15, 13kDa-A, and13kDa-B. The 2OkDa polypeptide was recognized as a novel component of IP for the first time. The 13kDa-A polypeptide was composed of 96 amino acids with a molecular weight of 10536. The 13kDa-B polypeptide consisted of 114 amino acids and had an acetylated amino terminus. The molecular weight of this protein was calculated to be 13130 including the acetyl group. The 13kDa polypeptides might be iron-sulfur proteins because the characteristic arrangement of cysteine residues were present in the 13kDa-A polypeptide. The partial amino acid sequences of the 30 and 20kDa polypeptides were also determined. Comparison of sequences revealed significant sequence similarities of the 30 and 20kDa polypeptides to the products of the ORF158 and psbG gene encoded in the chloroplast genome, respectively. The conserved sequence in the 2OkDa and PsbG proteins was also found in the small subunit of the nickel-containing hydrogenases. The conserved region among these proteins contained two cysteine residues, which might be involved in the formation of an iron-sulfur cluster. These results suggest that complex I is related to other redox enzyme complexes.

线粒体NADH：泛醌氧化还原酶（复合物I）是呼吸链中最复杂的系统。它由许多亚基组成，其中一些含有铁硫簇，但结构信息仍然有限。牛心脏复合物I的两个亲水亚组分通过层析系统地拆分成单个多肽。从复合物Ⅰ的黄素蛋白组分（FP）中分离出三种多肽（51、24和9 kDa），并测定了9 kDa多肽的完整氨基酸序列。该多肽分子量为9 kDa，由75个氨基酸组成，分子量为8437。该蛋白与其他蛋白没有明显的序列相似性。复合物Ⅰ的铁硫蛋白组分（IP）被分离成8个多肽，分别为75、49、30、20、18、15、13 kDa-A和13 kDa-B。20 kDa多肽首次被确认为IP的新组分。13 kDa-A多肽由96个氨基酸组成，分子量为10536。13 kDa-B多肽由114个氨基酸组成，具有乙酰化的氨基末端。该蛋白质的分子量计算为13130，包括乙酰基。13 kDa多肽可能是铁硫蛋白，因为13 kDa多肽中存在半胱氨酸残基的特征性排列。测定了30和20 kDa多肽的部分氨基酸序列。序列比较显示，30和20 kDa的多肽的显着序列相似性的ORF 158和psbG基因编码的叶绿体基因组中的产品，分别。在20 kDa和PsbG蛋白中的保守序列也被发现在含镍氢化酶的小亚基中。这些蛋白质的保守区含有两个半胱氨酸残基，这可能是参与铁硫簇的形成。这些结果表明复合物I与其他氧化还原酶复合物有关。

项目成果

Ryoji Masui: "The amino acid sequences of two 13kDa polypeptides and partial amino acid sequence of 30kDa polypeptide of complex I from bovine heart mitochondria:Possible location of iron-sulfur clusters" Journal of Biochemistry. 109. 534-543 (1991)

Ryoji Masui：“来自牛心脏线粒体的复合物 I 的两个 13kDa 多肽的氨基酸序列和 30kDa 多肽的部分氨基酸序列：铁硫簇的可能位置”《生物化学杂志》。

R.Masui: "The amino acid sequences of two 13kDa polypeptides and partial amino acid sequence of 30kDa polypeptide of complex I from bovine heart mitochondria" Journal of Biochemistry. 109. (1991)

R.Masui：“来自牛心脏线粒体的复合物 I 的两个 13kDa 多肽的氨基酸序列和 30kDa 多肽的部分氨基酸序列”《生物化学杂志》。

Sadao Wakabayashi,Ryoji Masui,Hiroshi Matsubara,and Youssef Hatefi (pp.49-55): "Bioenergetics.Molecular Biology,Biochemistry,and Pathology (Edited by Chong H.Kim and Takayuki Ozawa)" Plenum Press,New York, 1-484 (1990)

Sadao Wakabayashi、Ryoji Masui、Hiroshi Matsubara 和 Youssef Hatefi（第 49-55 页）：“生物能量学。分子生物学、生物化学和病理学（由 Chong H.Kim 和 Takayuki Ozawa 编辑）” Plenum Press，纽约，1-

R.Masui: "Amino acid sequences of two 13kDa polypeptides and partial amino acid sequence of 30kDa polypeptide of complex I from bovine Mt." The Protein Soc.symp.IV. M-125 (1990)

R.Masui：“来自牛 Mt. 的复合物 I 的两个 13kDa 多肽的氨基酸序列和 30kDa 多肽的部分氨基酸序列。”

Mutsuo Yamaguchi: "Mitochondrial energy-linked nicotinamide nucleotide transhydrogenase:Effect of substrates on the sensitivity of the enzyme to trypsin and identification of tryptic cleavage sites" Biochemistry. 29. 4136-4143 (1990)

Mutsuo Yamaguchi：“线粒体能量连接烟酰胺核苷酸转氢酶：底物对胰蛋白酶敏感性的影响以及胰蛋白酶切割位点的鉴定”生物化学。