Studies on Cyanide-Resistant Respiration in yeast, Hansenula anomala

异常汉逊酵母的抗氰呼吸作用的研究

• 批准号: 02680162
• 负责人: YOSHIMOTO Akio
• 金额: $ 1.28万
• 依托单位: Niigata College of Pharmacy
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02680162/
• 关键词: Hansenula anomala; Cyanide-resistant respiration; Superoxide anion radical; Radical scavengers; Role of Fe^<2+> in cyanide-resistant respiration; Unsaturated fatty acids; Cation ionophores; ス-パ-オキシドアニオン; 不飽和脂肪酸類; イオノフォア類; Qサイクル; フラボノイド類

(1) Mitochondria exihibiting cyanide-resistant respiration were isolated from Hansenula anomala incubated in the presence of antimycin A or a certain sulfur-compounds. Analysis of mitochondrial proteins by gel electrophoresis indicates that a 36 kDa protein was induced. When the cells were labeled with ^<35>S-methionine in the presence of both antimycin A and CCCP, the radioactivity was incorporated into a 39 kDa mitochondrial protein. Upon removal of CCCP, this protein was processed into a 36 kDa form, suggesting that the 39 kDa protein is the precursor of 36 kDa protein.(2) The antimycin A-dependent induction of cyanide-resistant respiration was completely repressed by specific chelators for Fe^<2+>. The addition of Fe^<2+> to the cells resulted in a rapid increase in cyanide-resistant respiration activity, suggesting that Fe^<2+> is involved in the reaction mechanism of this respiration.(3) Since a plant-like alternative oxidase was reported to serve as a sole terminal oxidase of Tr … More ypanosoma brucei brucei, the studies on the specific inhibitors of cyanide-resistant respiration have taken on a great sibnificance for the chemotherapy of trypanosome infections. We found specific inhibitory effects on cyanide-resistant respiration of some fatty acids with nonconjugated duoble bonds, radical scavengers such as CoQ derivatives and flavonoids, and cation ionophores.(4) Q_i site inhibitors such as antimycin A induce O_2 generation in respiring cyanide-sensitive mitochondria of H. anomala. Radical scavengers suppressed O_2 generation, and inhibited both the induction of cyanide-resistant respiration and the appearance of the mitochondrial 36 kDa protein. These results suggest that O_2 is involved in the induction of cyanide-resistant respiration.(5) A cDNA for mRNA induced by antimycin A was cloned. The nucleotide sequence revealed a long open reading frame of 342 codons encoding a protein. Immunoblotting showed that the 36 kDa protein localized in mitochondria was a mature form of the protein encoded by the cDNA. Less

(1)从异常汉逊酵母（Hansenula anomala）中分离出抗氰呼吸的线粒体。通过凝胶电泳分析线粒体蛋白表明诱导了一个36 kDa的蛋白。当细胞<35>在抗霉素A和CCCP同时存在的情况下用^S-甲硫氨酸标记时，放射性被掺入到39 kDa的线粒体蛋白中。去除CCCP后，该蛋白质被加工成36 kDa形式，表明39 kDa蛋白质是36 kDa蛋白质的前体。(2)抗氰呼吸的抗霉素A依赖性诱导被Fe^&lt;2+&gt;的特异性螯合剂完全抑制。向细胞中加入Fe^&lt;2+&gt;导致抗氰呼吸活性迅速增加，表明Fe^&lt;2+&gt;参与了这种呼吸的反应机制。(3)由于一种类似植物的替代氧化酶被报道作为Tr的唯一末端氧化酶， ...更多信息 布氏锥虫（ypanosoma brucei）的抗氰呼吸抑制剂的研究对锥虫感染的化疗具有重要意义。我们发现一些具有非共轭双键的脂肪酸，自由基清除剂如辅酶Q衍生物和黄酮类化合物，以及阳离子载体对抗氰呼吸有特定的抑制作用。(4)Q_i位点抑制剂如抗霉素A可诱导H.异常自由基清除剂抑制O_2的产生，抑制抗氰呼吸的诱导和线粒体36 kDa蛋白的出现。这些结果表明O_2参与了抗氰呼吸的诱导。(5)克隆了抗霉素A诱导的mRNA的cDNA。核苷酸序列揭示了一个长的开放阅读框架的342个密码子编码的蛋白质。免疫印迹显示，定位于线粒体中的36 kDa蛋白是由cDNA编码的蛋白的成熟形式。少

项目成果

N.Minagawa,S.Sakajo,and A.Yoshimoto: "Effects of Ionophores on CyanideーResistant Respiration of the Mitochondria Isolated from Hansenula anomala"

N. Minakawa、S. Sakajo 和 A. Yoshimoto：“离子载体对从异常汉逊酵母中分离出的线粒体的抗氰呼吸作用的影响”

N.Minagawa,K.kaneko,S.Sakajo,and A.Yoshimoto: "Effects of 8ーHydroxyquinoline and Related Compounds on CyanideーResistant Respiration of the Isolated Mitochondria from Hansenula anomala"

N. Minakawa、K. kaneko、S. Sakajo 和 A. Yoshimoto：“8-羟基喹啉和相关化合物对异常汉逊酵母分离线粒体的抗氰呼吸作用的影响”

N.Minagawa,S.Sakajo,T.Komiyama and A.Yoshimoto: "Essential Role of Ferrous Iron in Cyanide-Resistant Respiration in Hansenula anomala" FEBS Lett.267. 114-116 (1990)

N.Minakawa、S.Sakajo、T.Komiyama 和 A.Yoshimoto：“亚铁在异常汉逊酵母抗氰呼吸中的重要作用”FEBS Lett.267。

S.Sakajo,N,Minagawa,T.Komiyama,and A.Yoshimoto: "Molecular Cloning of cDNA for Antimycin AーInducible mRNA and Its Role in CyanideーResistant Respiration in Hansenula anomala" Biochim.Biophys.Acta. 1090. 102-108 (1991)

S. Sakajo、N、皆川、T. Komiyama 和 A. Yoshimoto：“抗霉素 A 诱导 mRNA 的 cDNA 分子克隆及其在异常汉逊酵母抗氰化物呼吸中的作用”Biochim.Biophys.Acta 1090。 108 (1991)

N. Minagawa, T. Takatsu, S. Sakajo, T. Komiyama and A. Yoshimoto: "Effects of Idebenone on Cyanide-Resistant Respiration of the Mitochondria Isolated from Hansenula anomala" Agric. Biol. Chem.55. 845-846 (1991)

N. Minakawa、T. Takatsu、S. Sakajo、T. Komiyama 和 A. Yoshimoto：“艾地苯醌对从异常汉逊酵母中分离出的线粒体抗氰化呼吸的影响”Agric。