Physiological function of novel Ascaris cytochrome b5 in adaptation to low-oxygen tension
新型蛔虫细胞色素b5适应低氧张力的生理功能
基本信息
- 批准号:12670241
- 负责人:
- 金额:$ 2.18万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:2000
- 资助国家:日本
- 起止时间:2000 至 2001
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
In the current research project, the novel cytochrome b5, previously demonstrated in Ascaris suum body wall, has been studied to elucidate its biosynthesis, distribution in the nematode's tissues and organella, and physiological functions. Immunoblotting analysis of subcellular fractions of the body wall and perienteric fluid suggested that A. suum cytochrome b5 was not localized in mitochondria nor in microsome, but distributed in cytosolic fraction, and secreted into the perienteric fluid. The clear and distinct staining was observed in the cuticle layers of the body wall in immunohistochemical studies, indicating that purified cytochrome b5 was derived from the cuticle layers. In contrast to cytosolic distribution of the cytochrome b5, enzymes catalyzing its reduction, i. e. NAD(P)H- cytochrome b5 reductase, were detected in mitochondrial and microsomal fractions, but not in the cytosolic fraction. The reductase activity was also present in the perienteric fluid. To study possible r … More oles of the presequence in biosynthesis or distribution, the gene encoding A. suum cytochrome b5 precursor protein, comprising a full-length presequence consisting of 30 amino acid residues and mature protein, and two genes encoding the cytochrome b5 with truncated presequence, one lacking 18, and the other lacking 28 amino acid residues from its N-terminus, were expressed in E. coli. Among them, only the cells transformed with cDNA encoding intact precursor protein were shown to produce processed cytochrome b5, which is indistinguishable from native protein purified from the body wall. The processed cytochrome b5 was demonstrated by subcellular fractionation to be transported to the bacterial periplasm, indicating that the nematode's presequence was functional in E. coli. Cytochrome b5 was partially purified from perienteric fluid of A. suum ; the spectral and sequence analysis showed that the nematode's perienteric fluid in deed contained functional cytochrome b5, not apo-protein lacking heme moiety. The cytochrome b5 was shown to be involved in methemoglobin reductase system in the body fluid. Bioinformatic studies showed that C. elegans possessed four homologues of cytochrome b5. Less
在当前的研究项目中,对先前在猪蛔虫体壁中展示的新型细胞色素b5进行了研究,以阐明其生物合成、在线虫组织和细胞器中的分布,以及体壁和肠周亚细胞部分的免疫印迹分析。液体表明 A. suum 细胞色素 b5 既不定位于线粒体也不定位于微粒体,而是分布于胞质部分,并分泌到肠周免疫组织化学研究中在体壁的角质层中观察到清晰且明显的染色,表明纯化的细胞色素b5源自角质层,与细胞色素b5的细胞质分布相反,酶催化其还原,即NAD。在线粒体和微粒体部分中检测到 (P)H-细胞色素 b5 还原酶,但在胞质部分中未检测到。肠周中也存在还原酶活性。为了研究前序列在生物合成或分布中可能的作用,编码猪细胞色素b5前体蛋白的基因,包括由30个氨基酸残基和成熟蛋白组成的全长前序列,以及编码细胞色素的两个基因。其中,前序列b5的N端缺失18个氨基酸残基,另一种缺失28个氨基酸残基,其中,仅在大肠杆菌中表达。用编码完整前体蛋白的 cDNA 转化的细胞显示出产生加工的细胞色素 b5,其与从体壁纯化的天然蛋白没有区别。通过亚细胞分级分离证明加工的细胞色素 b5 被转运到细菌周质,这表明线虫的细胞色素 b5前序列在大肠杆菌中是有功能的。肠周液中确实含有功能性细胞色素b5,而不是缺乏血红素部分的载脂蛋白。生物信息学研究表明,线虫具有四种细胞色素b5的同源物。
项目成果
期刊论文数量(6)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Amino, H.: "Stage-specific isoforms of Ascaris suum complex II : the fumarate reductase of the parasitic adult and the succinate dehydrogenase of free-living larvae share a common iron-sulfur subunit"Mol.Biochem.Parasitol. 106. 63-76 (2000)
Amino, H.:“猪蛔虫复合体 II 的阶段特异性亚型:寄生成虫的富马酸还原酶和自由生活幼虫的琥珀酸脱氢酶共享一个共同的铁硫亚基”Mol.Biochem.Parasitol。
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- 影响因子:0
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- 通讯作者:
Amino, H., Wang, H., Hirawake, H., Saruta, F., Mizuchi, D., Mineki, R., Shindo, N., Murayama, K., Takamiya, S., Aoki, T., Kojima, S., and Kita, K.: "Stage-specific isoforms of Ascaris suum complex II : the fumarate reductase of the parasitic adult and the
Amino, H.、Wang, H.、Hirawake, H.、Saruta, F.、Mizuchi, D.、Mineki, R.、Shindo, N.、Murayama, K.、Takamiya, S.、Aoki, T.、
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- 影响因子:0
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Kita, K.: "Electron transfer Complexes in Ascaris Mitochondria"Adv.Parasitol. (in press).
Kita, K.:“蛔虫线粒体中的电子转移复合物”Adv.Parasitol。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Amino, H.: "Stage-specific isoforms of Ascaris suum complex II : the fumarate reductase of the parasitic adult and the succinate dehydrogenase of free-living larvae share a common iron-sulfur subunit"Mol. Biochem. Parasitol.. 106. 63-76 (2000)
Amino,H.:“猪蛔虫复合体 II 的阶段特异性亚型:寄生成虫的富马酸还原酶和自由生活幼虫的琥珀酸脱氢酶共享一个共同的铁硫亚基”Mol。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Kita, K., and Takamiya, s.: "Electron transfer Complexes in Ascaris Mitochondria."Adv. Parasitol. (in press).
Kita, K. 和 Takamiya, s.:“蛔虫线粒体中的电子转移复合物”。
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- 影响因子:0
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TAKAMIYA Shinzaburo其他文献
TAKAMIYA Shinzaburo的其他文献
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{{ truncateString('TAKAMIYA Shinzaburo', 18)}}的其他基金
Proteomic analyses of Ascaris mitochondrial respiratory chain
蛔虫线粒体呼吸链的蛋白质组学分析
- 批准号:
22590383 - 财政年份:2010
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Structural and functional analysis of Ascaris suum cytochrome b in the nematode methemoglobin reductase system
线虫高铁血红蛋白还原酶系统中猪蛔虫细胞色素b的结构和功能分析
- 批准号:
18590406 - 财政年份:2006
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Proteomic analyses of oxygen -responding proteins of Ascaris suum nematodes
猪蛔虫线虫氧响应蛋白的蛋白质组学分析
- 批准号:
14570220 - 财政年份:2002
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Molecular properties of cytochrome c oxidase in Ascaris respiratory chain and its response to oxygen tension
蛔虫呼吸链细胞色素c氧化酶的分子特性及其对氧张力的响应
- 批准号:
10670239 - 财政年份:1998
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Biochemical studies on aerobic-anaerobic respiratory transition in mitochondria from parasitic helminthes
寄生蠕虫线粒体有氧-无氧呼吸转变的生化研究
- 批准号:
03670201 - 财政年份:1991
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
Changes and Their Control Mechanisms of Mitochondrial Electron-Transport Components During Ascaris Life Cycle
蛔虫生命周期线粒体电子传递成分的变化及其控制机制
- 批准号:
01570223 - 财政年份:1989
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
相似海外基金
Structural and functional analysis of Ascaris suum cytochrome b in the nematode methemoglobin reductase system
线虫高铁血红蛋白还原酶系统中猪蛔虫细胞色素b的结构和功能分析
- 批准号:
18590406 - 财政年份:2006
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Proteomic analyses of oxygen -responding proteins of Ascaris suum nematodes
猪蛔虫线虫氧响应蛋白的蛋白质组学分析
- 批准号:
14570220 - 财政年份:2002
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Physiological function of respiratory enzymes specific for parasite mitochondria in the adaptation to low oxygen tension
寄生虫线粒体特异性呼吸酶在适应低氧张力中的生理功能
- 批准号:
13854011 - 财政年份:2001
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for Scientific Research (S)