Elucidation of Reaction Mechanisms of Biological Nitric Oxide by Vibrational Spectroscopy

振动光谱法阐明生物一氧化氮的反应机制

• 批准号: 07454157
• 负责人: KITAGAWA Teizo
• 金额: $ 5.06万
• 依托单位: (Institute for Molecular Science) Okazaki National Research Institutes
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07454157/
• 关键词: Nitric Oxide; Guanylate Cyclase; Resonance Raman; Vibrational Spectroscopy; NO stretching mode; 赤外吸収; ダイオードレーザー; 振動分光法; 共鳴ラマン散乱

Soluble guanylate cyclase (sGC) was isolated from bovine lung and its resonance Raman (RR) spectra were investigated for the reduced, CO-bound (CO-sGC), NO-bound (NO-sGC), oxidized, and oxidized NO-bound forms in the presence and absence of GTP.The RR spectra of sGC and CO-sGC,including the Fe-His stretch at 204 cm^<-1> and Fe-CO stretch at 473 cm^<-1>, were unaltered by binding of GTP,but apparent RR spectra of NO-sGC in the presence of GTP changed with time and concentrations of GTP.In the absence of GTP,the RR bands of N-O stretch (nu_<NO>) and Fe-No stretch (nu_<Fe-NO>) were observed at 1679 and 521 cm^<-1>, respectively. In its presence, two types of RR spectra were obtained, but after all GTP was exhausted by the enzymatic reaction, the spectrum for the absence of GTP was restoed. In one type of RR spectra, which appeared prior to the other, no NO-isotope sensitive band was detected and is heme spectrum was of a five-coordinate ferric high-spin type. In the other type, two nu_<NO> bands were observed at 1700 and 1681 cm^<-1>, which exhibited ^<15>NO isotopic frequency shifts of 21 and 34 cm^<-1>, respectively. Structural implications of the two types of RR spectra arizing from reaction intermediates have been discussed.

从牛肺中提取了可溶性鸟苷环化酶(SGC)，研究了在GTP存在和不存在的情况下，还原的、CO结合的(CO-sGC)、非结合的(NO-sGC)、氧化的和氧化的非结合形式的共振拉曼光谱。，但在GTP存在下，NO-sGC的表观RR谱随时间和浓度的变化而变化。在没有GTP的情况下，N-O伸展(nu&lt；NO&gt；)和Fe-非伸展(nu&lt；Fe-NO&gt；)的RR谱带分别出现在1679和521 cm^&lt；-1&gt；在其存在下，可获得两种类型的RR光谱，但在酶反应耗尽所有GTP后，恢复了不含GTP的光谱。在一种类型的RR谱中，没有检测到非同位素敏感带，并且在另一种类型的RR谱中，它是一个五配位铁的高自旋型光谱。在另一种类型中，在1700和1681 cm；-1&gt；处观察到两条nu&lt；no&gt；带，它们分别表现出21和34 cm^&lt；-1&gt；无同位素频移。讨论了反应中间体产生的两种RR谱的结构含义。

项目成果

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P.Jewsbury：“碳单氧肌红蛋白中的远端 CO 相互作用：3 个远端突变体的分子动力学。”

T.Kohzuma: "Spectroscopic and electrochemical studies on active-site transitions of the type 1 copper protein pseudoazurin from Achromobacter cycloclastes." J.Biol.Cham.270. 25733-25738 (1995)

T.Kohzuma：“来自环碎无色杆菌的 1 型铜蛋白假天青蛋白活性位点转变的光谱和电化学研究。”

S.Hirota: "Observation of nonfundamental Fe-O_2 and Fe-CO vibrations and potential anharmonicities for oxy-and carbonmonoxy-hemoglobin ; Evidence supporting a new assignment of the Fe-C-O bending fundamental." J.Am.Chem.Soc.117. 821-822 (1995)

S.Hirota：“观察非基本 Fe-O_2 和 Fe-CO 振动以及氧和碳单氧血红蛋白的潜在非谐性；支持 Fe-C-O 弯曲基本原理新分配的证据。”

D.A.Proshlyakov: "Microcirculating system for simultaneous determination of Raman and absorption spectra of enzymatic reaction intermediates and its application to the reaction of cytochrome c oxidase with hydrogen peroxide." Biochemistry. 35. 76-82 (1996

D.A.Proshlyakov：“同时测定酶反应中间体拉曼光谱和吸收光谱的微循环系统及其在细胞色素 C 氧化酶与过氧化氢反应中的应用。”

M.Nagai: "Ultraviolet resonance Raman studies of quaternary structure of hemoglobin using a tryptophan b37 mutant." J.Biol.Chem.270. 1636-1642 (1995)

M.Nagai：“使用色氨酸 b37 突变体对血红蛋白四级结构进行紫外共振拉曼研究。”