SYNTHETIC MODELING OF COPPER PROTEIN ACTIVE SITES

铜蛋白活性位点的综合建模

• 批准号: 6192240
• 负责人: WILLIAM B Tolman
• 金额: $ 24.6万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-08-01 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6192240
• 关键词: Raman spectrometry; active sites; chemical models; chemical synthesis; copper; cytochrome oxidase; electron spin resonance spectroscopy; electron transport; hydrogen bond; metalloproteins; model design /development; nitric oxide; nitrous oxide; oxygen; oxygenases

DESCRIPTION: (adapted from applicant's abstract) Proteins that contain copper in their active sites represent a large and functionally significant class of metallobiomolecules that play central roles in life processes. Electro transfer, reversible binding and activation of dioxygen, oxidation of organic molecules, and nitrogen oxide activation are illustrative examples of the wide range of important reactions performed by copper proteins. This functional diversity is matched by a high degree of variability in the geometric, electronic structural and spectroscopic features of the copper active sites. Despite extensive efforts to relate these features to functional attributes through experimental and theoretical studies, our understanding of structure/function relationships at the molecular level is incomplete and many questions concerning the detailed mechanisms of copper-mediated processes in biology remain unanswered. Such issues will be addressed in the proposed research through the synthetic modeling approach, wherein low molecular weight complexes designed to replicate metalloprotein active site structure and function are characterized and their reactivity examined. Through this approach, fundamental chemical insights into copper site structure and mechanisms of action will be obtained. The specific aims of the proposed research are to: 1.Understand the mechanisms of nitrogen oxide processing by copper proteins such as nitrite and nitrous oxide reductase, important players in the global nitrogen cycle. Toward this end, the synthesis , characterization, and reactivity of copper-hyponitrite complexes will be targeted. 2.Understand how structural perturbations influence the function of the ubiquitous copper-thiolate electron transfer sites. In particular, the synthesis and characterization of models of perturbed mononuclear type 1 and dinuclear, mixed-valence CuA centers will be pursued. 3.Obtain fundamental chemical information on the pathways of dioxygen activation at mononuclear copper sites such as those found in dopamine-beta-monooxygenase and peptidylglycine alpha hydroxylating monooxygenase, important catalysts in mammalian hormone biosynthesis. The synthesis, characterization , and reactivity of new monocopper-dioxygen species will be studied. 4.Understand dioxygen activation at trinuclear copper sites found in the multicopper oxidases and particulate methane monooxygenase by targeting the synthesis and O2 reactivity of tricopper(I) complexes of preorganized N-donor ligands. 5.Provide new mechanistic insights into copper-mediated cofactor biogenesis in the amine oxidases and galactose oxidase, both of which have unusual organic cofactors that are post-translationally modified in reactions involving Cu and O2 . In particular, studies of the activation of coordinated phenolate and/or thiophenolate ligands will be performed.

描述：（改编自申请人的摘要）含有铜的蛋白质 在其主动地点中，代表了大型且功能意义的类别 在人生过程中起着核心作用的金属生物分子。电 二氧化物的转移，可逆结合和激活，有机的氧化 分子和氮氧化物激活是宽的例子 铜蛋白执行的重要反应范围。这个功能 多样性与几何可变性相匹配， 铜主动部位的电子结构和光谱特征。 尽管大力将这些功能与功能属性联系起来 通过实验和理论研究，我们对 分子水平的结构/功能关系不完整，许多 有关铜介导过程的详细机制的问题 生物学仍未得到答复。这些问题将在拟议的 通过合成建模方法进行研究，其中低分子量 旨在复制金属蛋白活性位点结构的复合物和 功能是表征的，其反应性检查。通过这个 方法，对铜现场结构的基本化学见解和 将获得作用机制。提议的具体目的 研究是： 1.理解铜蛋白的氮氧化物加工机制 例如亚硝酸盐和一氧化二氮还原酶，全球重要参与者 氮循环。为此，综合，表征和 铜 - 高硝酸盐配合物的反应性将被靶向。 2.理解结构扰动如何影响 普遍存在的铜硫酸硫酸电子转移位点。特别是 扰动单核类型1和的模型的合成和表征 将追求围栏，混合价CUA中心。 3.关于二氧化物途径的Obtain基本化学信息 在单核铜部位的激活，例如在 多巴胺-beta-偶加酶和肽基甘氨酸α-羟基羟基化 单加氧酶，哺乳动物激素生物合成中的重要催化剂。这 新单室二氧化物的合成，表征和反应性 将被研究。 4.理解在三核铜部位的二氧化激活 通过靶向多磷酸氧化酶和颗粒甲烷单加氧酶 三角体的合成和O2反应性（i）复合物的n-donor复合物 配体。 5.对铜介导的辅因子生物发生的新机械见解 胺氧化酶和半乳糖氧化酶都有不同寻常的有机物 在涉及Cu和的反应中经过翻译后修改的辅助因子 O2。特别是，对协调苯酸酯和/或的激活的研究 将进行硫芬酸酯配体。