Ultraviolet Resonance Raman Study on Protein Structure Dynamics of Cytochrome c Oxidase

细胞色素c氧化酶蛋白质结构动力学的紫外共振拉曼研究

• 批准号: 08680730
• 负责人: OGURA Takashi
• 金额: $ 1.34万
• 依托单位: Okazaki National Research Institutes
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08680730/
• 关键词: cytochrome c oxidase; UV resonance Raman; bioenergetics; proton pump; oxygen activation; electron transfer

A high performance ultraviolet resonance Raman spectrophotometer for biological applications has been constructed. A solar-blind intensified charge-coupled device was attached to a 1260 mm single spectrograph with a 200 nm-blazed, 3600 grooves/mm grating. The 488 nm output of a mode-lacked argon ion laser was frequency doubled to obtain 244nm UV light for resonance Raman excitation. This Raman spectrophotometer has been applied to cytochrome c oxidase to probe structural changes of tyrosine and tryptophan residues upon redox and ligation-state changes. Resting oxidized, fully-reduced and mixed-valence forms of the enzyme in the presence and absence of carbonmonoxide or cyanide have been measured. It became evident that a redox change of heme a and / or Cu_A induced most prominent changes on tyrosine and tryptophan residues while ligation to heme a_3 and redox change of heme a_3 and / or Cu_B less affected the protein structures. Basic data to analyze time-resolved UV resonance Raman spectra of cytochrome c oxidase have been established.

构建了一种用于生物应用的高性能紫外共振拉曼分光光度计。将日盲增强电荷耦合装置连接到带有 200 nm 闪耀、3600 槽/mm 光栅的 1260 mm 单光谱仪上。将无模氩离子激光器的 488 nm 输出倍频以获得用于共振拉曼激发的 244 nm 紫外光。该拉曼分光光度计已应用于细胞色素 c 氧化酶，以探测氧化还原和连接状态变化时酪氨酸和色氨酸残基的结构变化。在存在和不存在一氧化碳或氰化物的情况下，测量了酶的静止氧化、完全还原和混合价形式。很明显，血红素a和/或Cu_A的氧化还原变化引起酪氨酸和色氨酸残基的最显着变化，而与血红素a_3的连接以及血红素a_3和/或Cu_B的氧化还原变化对蛋白质结构的影响较小。分析细胞色素c氧化酶时间分辨紫外共振拉曼光谱的基本数据已经建立。

项目成果

Teizo Kitagawa and Takashi Ogura: Progress in Inorganic Chemistry, Vol.45. John Wiley and Sons, Inc., 49 (1997)

Teizo Kitakawa 和 Takashi Ogura：无机化学进展，第 45 卷。

T.Kitagawa and T.Ogura: "Progress in Inorganic Chemistry,vol.45 (K.D.karlin ed.)" John Wiley & Sons,Inc., 49 (1997)

T.Kitakawa 和 T.Ogura：“无机化学进展，第 45 卷（K.D.karlin 编辑）”John Wiley

T.Ogura,S.Hirota,D.A.Proshlyakov et al.: "Time-Resolved Resonance Raman Evidence for Tight Coupling between Electron Transfer and Proton Pumping of Cytochrome c Oxidase...." J.Am.Chem.Soc.118. 5443-5449 (1996)

T.Ogura、S.Hirota、D.A.Proshlyakov 等人：“细胞色素 c 氧化酶的电子转移和质子泵送之间紧密耦合的时间分辨共振拉曼证据......”J.Am.Chem.Soc.118。

Masahiro Mukai: "Effects of Concerted Hydrogen Bonding of Distal Histidine on Active Site Structures of Horseradish Peroxidase:Resonance...." J.Am.Chem.Soc.119. 1758-1766 (1997)

Masahiro Mukai：“远端组氨酸协同氢键对辣根过氧化物酶活性位点结构的影响：共振……”J.Am.Chem.Soc.119。

Xiaojie Zhao: "Observation of the Resonance Raman Spectra of the Q_A^<.-> and Q_B^<.-> in photosynthetic Reaction Center from sphaeroides R26" J.Am.Chem.Soc.119. 5263-5264 (1997)

赵晓杰：“球状体R26光合反应中心Q_A^<.->和Q_B^<.->共振拉曼光谱的观察”J.Am.Chem.Soc.119。