Development of a fast scanning IR spectrophotmeter and its application to oxygenase systems.
快速扫描红外分光光度计的开发及其在加氧酶系统中的应用。
基本信息
- 批准号:59880012
- 负责人:
- 金额:$ 6.21万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Developmental Scientific Research
- 财政年份:1984
- 资助国家:日本
- 起止时间:1984 至 1985
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Aerobic organisms utilize molecular oxygen ( <O_2> ) in many ways. Such oxygen utilizing reactions include various oxidase and oxygenase reactions, and superoxide-producing reactions. Under physiologic conditions, however, molecular oxygen is mainly in an inactive triplet groud state which usaully requires an "activation" to react with singlet organic molecules. Thus, the mechanisms for activation of molecular oxygen has been one of the main themr in biochrmistry. In the present study, we developed a new type of IR spectrophotometer which enables us to measure O-O stretching frequency around 1100 <cm^(-1)> of molecular oxygen bound to iron in heme-containing oxygenases and oxidases. Since such oxygen complexes are all unstable reaction intermediates, it requires fast scanning together with high sensitivity, which are not fullfilled by any dispersive and/or FT type of IR spectrophotometer. For this purpose, we succesfully developed a new type of machine by employing MCT, high frequency chopper and high speed scanning device with special soft wear systems. The spectrophotmeter system made us possible to measure the O-O stretching of oxygen in oxy-CoMb (4 mM) within 7 minutes at 10゜C. In preliminary experiments, we also succeeded in measuring the O-O stretching of oxy-P450cam which was stabilized by utilizing the diacetylheme-substituted enzyme.
好氧生物以多种方式利用分子氧<O_2>。这种利用氧的反应包括各种氧化酶和加氧酶反应,以及产生超氧化物的反应。然而,在生理条件下,分子氧主要处于无活性的三重基态,通常需要“活化”才能与单线态有机分子反应。因此,分子氧的活化机制一直是生物化学研究的热点之一。本研究中,我们研制了一种新型的红外分光光度计,使我们能够测量含血红素的加氧酶和氧化酶中与铁结合的分子氧在1100 cm^(-1)附近的O-O伸缩频率。由于这些氧络合物都是不稳定的反应中间体,因此需要快速扫描以及高灵敏度,这是任何色散型和/或FT型红外分光光度计所不能满足的。为此,我们成功地开发了一种新型的机器,采用MCT,高频斩波器和高速扫描装置与特殊的软磨损系统。荧光计系统使我们能够在10 ℃下7分钟内测量oxy-CoMb(4 mM)中氧的O-O伸缩。在初步实验中,我们还成功地测量了氧-P450 cam的O-O伸缩,该氧-P450 cam通过利用二乙酰血红素取代的酶而稳定。
项目成果
期刊论文数量(8)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Proceedings of the 7th International Congress on Raman spectroscopy. p.492-493 (1984)
第七届国际拉曼光谱大会论文集。
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ISHIMURA Yuzuru其他文献
ISHIMURA Yuzuru的其他文献
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{{ truncateString('ISHIMURA Yuzuru', 18)}}的其他基金
MECHANISM FOR OXYGEN ACTIVATION BY CYTOCHROME P450cam
CYTOCHROME P450cam 的氧气激活机制
- 批准号:
07458163 - 财政年份:1995
- 资助金额:
$ 6.21万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Oxygen activation mechanisms in cytochrome P450-and NADPH oxidase-catalyzed reactions
细胞色素 P450 和 NADPH 氧化酶催化反应中的氧活化机制
- 批准号:
05454631 - 财政年份:1993
- 资助金额:
$ 6.21万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)
Purification and Molecular Mechanisms of O^-_ generating System in Polymorphonuclear Leukocytes and Thyroid Cells
多形核白细胞和甲状腺细胞O^-_生成系统的纯化及分子机制
- 批准号:
63480505 - 财政年份:1988
- 资助金额:
$ 6.21万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)
Studies on electronic structures and reactivities of an oxygenated form of heme-containing enzymes by vibrational spectroscopic methods
通过振动光谱法研究含氧形式的血红素酶的电子结构和反应性
- 批准号:
61480469 - 财政年份:1986
- 资助金额:
$ 6.21万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)
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