CYRO- & TIME-RESOLVED VIBRATION STUDY OF METALLOPROTEINS

氧化还原金属蛋白是所有生物生命过程所必需的 有机体。氧化还原活性中心的基础研究 金属蛋白，包括从相对复杂到相对复杂的系统 提出了将简单的电子转移蛋白转化为“多头”氧化酶。 我们的目标是理解，在基本的物理和 金属位置的化学性质及其与金属离子的相互作用 蛋白质结构，这些蛋白质是如何完成其易化功能的 电子转移和氧等小分子的活化。 鉴于大量的病理性疾病，这种理解是至关重要的 与氧化还原金属蛋白功能有关的条件。 提出的主要实验方法是振动光谱学， 包括共振拉曼光谱(RR)和傅里叶变换红外光谱(FTIR)。 重点是：RR和FTIR研究作为温度的函数 低温和环境条件之间的关系；以及时间分辨振动 光谱学(共振拉曼TR3，FTIR，TR-FTIR) 配位和电子转移动力学。最初的目标系统是 简单的铜“蓝”蛋白，例如，藻蓝蛋白、天青素和 木犀草素；多铜氧化酶，如漆酶，抗坏血酸 酶、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。预期结果包括： 提高光谱分辨率和信噪比，从而实现更可靠 传统意义上的振动数据分析；来自 频率和线形的温度依赖关系，关于 基本振动动力学及其对结构的可能影响 电子转移过程；以及可能观测到的极低能量 第三类中的电子跃迁，如自旋-自旋相互作用 铜对或在细胞色素A3中的细胞色素氧化酶。TR3和TR-FTIR 研究针对多铜氧化酶和细胞色素氧化酶。 预期结果包括：阐明瞬变的结构 酶在所有时间尺度上的功能；02之前的结合模式 细胞色素氧化酶中血红素的氧化和自旋状态 以及蛋白质约束对暂态结构的影响 金属遗址。给出了来自低温装置的改进的振动数据 研究，包括同位素地点。给出了来自 低温研究，包括同位素数据，现代技术的应用 金属蛋白活性的小分子振动分析方法 建议选址。实验结果及其分析将 导致可靠的结构、动力学和功能解释 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.