CYRO- & TIME-RESOLVED VIBRATION STUDY OF METALLOPROTEINS

氧化还原金属蛋白是所有生命过程所必需的 有机体 氧化还原活性中心的基础研究 金属蛋白，包括复杂程度从相对 简单的电子转移蛋白质到“多头”氧化酶。 目标是了解，在基本的物理和 金属位点的化学性质及其与 蛋白质结构，这些蛋白质如何完成它们的功能， 电子转移和小分子如氧的活化。 这种理解是至关重要的，因为大量的病理性 与氧化还原金属蛋白功能相关的条件。 提出的主要实验方法是振动光谱， 包括共振拉曼（RR）和傅里叶变换红外（FTIR）。 重点放在：RR和FTIR研究作为温度的函数 在低温和环境条件之间;和时间分辨的振动 光谱学（共振拉曼TR 3、FTIR、TR-FTIR） 连接和电子转移动力学。 最初的目标系统是 简单的铜“蓝”蛋白，例如，质体蓝素，天青蛋白，和 多铜氧化酶，例如漆酶、抗坏血酸 氧化酶和铜蓝蛋白;和细胞色素氧化酶。 所有这些系统 将通过低温振动技术进行研究。 预期成果包括： 提高了光谱分辨率和信噪比， 传统意义上的振动数据分析;从 频率和线型的温度依赖性，有关信息 基本的振动动力学及其可能的影响 电子转移过程;以及可能观测到极低能量 电子跃迁，如3型中的自旋-自旋相互作用 铜对或在细胞色素氧化酶的细胞色素a3中。 TR 3和TR-FTIR 研究针对多铜氧化酶和细胞色素氧化酶。 预期的结果包括： 酶在所有时间尺度上的功能; 02的还原;细胞色素氧化酶中血红素的氧化态和自旋态 瞬时;以及蛋白质限制对瞬时结构的影响 金属网站 从低温实验中得到的振动数据 研究，包括同位素场址。 鉴于改进的振动数据， 低温研究，包括同位素数据，现代 金属蛋白活性的小分子振动分析方法 网站已提出。 实验结果及其分析将 导致可靠的结构，动力学和功能的解释， 振动数据

The redox metalloproteins are necessary for the life processes of all organisms. Fundamental studies of the active sites of redox metalloproteins, including systems ranging in complexity from relatively simple electron-transfer proteins to "multiheaded" oxidases, are proposed. The objectives are to understand, in terms of the fundamental physical and chemical properties of the metal sites and their interactions with the protein structure, how these proteins accomplish their functions of facile electron transfer and the activation of small molecules such as oxygen. Such understanding is crucial in view of the large number of pathological conditions which are related to the function of the redox metalloproteins. The primary experimental approach proposed is vibrational spectroscopy, including resonance Raman (RR) and Fourier transform infrared (FTIR). Emphasis is placed upon: RR and FTIR studies as a function of temperature between cryogenic and ambient conditions; and time-resolved vibrational spectroscopy (resonance Raman TR3, FTIR, TR-FTIR) of transients in the ligation and electron transfer dynamics. The initial target systems are the simple copper "blue" proteins, e.g., plastocyanin, azurin, and stallacyanin; the multicopper oxidases, e.g. the laccases, ascorbate oxidase, and ceruloplasmin; and cytochrome oxidase. All of these systems will be studied by cryo-vibrational techniques. Expected results include: improved spectral resolution and signal-to-noise, leading to more reliable analysis of the vibrational data in the conventional sense; from the temperature dependence of frequencies and line-shapes, information on fundamental vibrational dynamics and their possible influence on the electron transfer process; and possible observation of very low energy electronic transitions such as the spin-spin interactions in the type 3 copper pair or in cytochrome a3 of cytochrome oxidase. The TR3 and TR-FTIR studies are directed at the multi-copper oxidases and cytochrome oxidase. Expected results include: elucidation of the structures of transients in the function of the enzymes at all timescales; modes of 02 binding before reduction of 02; oxidation and spin states of hemes in cytochrome oxidase transients; and effects of protein constraints on transient structures of the metal sites. Given improved vibrational data from the cryogenic studies, including isotope sites. Given improved vibrational data from the cryogenic studies, including isotope data, application of modern small-molecule vibrational analysis methods to the metalloprotein active sites is proposed. The experimental results and the analysis thereof will lead to reliable structural, dynamical and functional interpretations of vibrational data.