SYNTHESES OF LABELED HEMES FOR PROTEIN NMR STUDIES

建议开发和利用方法，使 合成预定区域选择性标记的天然血红素 具有氘、碳13、氚、氮15和氟的位置， 这些将用作核磁共振和共振拉曼探针以获得 了解血红素载脂蛋白相互作用和结构/功能 多种具有生物学重要意义的血红素蛋白之间的关系，例如 血红蛋白、肌红蛋白、细胞色素和过氧化物酶。 还建议 合成各种未标记的血红素类似物、血红素二聚体、铁 氧代绿素和氢血红素。 拟议的工作将为 了解血红素蛋白功能、结构和电子 导致贫血等多种衰弱性疾病的因素。 的 合成化合物还将用于 EPR、X 射线、MCD 和其他 与许多独立的机构合作进行生物学研究 调查人员。 重建为适当的脱辅基蛋白后， 新型合成化合物将用于生产光谱 分配，以及简化光谱和解释 多种血红素蛋白。 两种基本的合成方法 将开发新化合物；碳13、氮15、氟 标记的和未标记的卟啉将通过全合成获得 无环前体。 此类方法的方法已经被研究出来 已出炉，但已计划改进和发现新方法 预计。 一些氘标记、碳13标记的卟啉， 原卟啉 IX 类似物、氧代绿素、磺基血红素和其他二氢卟酚 血红素将通过操纵现有的取代基来接近 市售的卟啉或来自叶绿素 α 的 从市售藻类中提取。 经过方法发现和 优化，并为了充分利用综合发展 通常只在有限的范围内进行，我们计划准备大部分 批量标记、未标记和结构修饰的血红素 约100毫克；然后将这些化合物提供给 合作者使生物学上重要的多核核磁共振、共振 重建血红素的拉曼、EPR、X 射线、MCD 和其他特性 血红素蛋白、非铁金属复合物及其结构修饰 卟啉配体可以被研究、关联、合理化和 解释了。

It is proposed to develop and exploit methodology which will permit synthesis of natural hemes regioselectively labeled in predetermined positions with deuterium, carbon- 13, tritium, nitrogen- 15, and fluorine, and these will be used as NMR and resonance Raman probes to gain an understanding of heme apoprotein interactions and structure/function relationships in a variety of biologically important heme proteins such as hemoglobins, myoglobins, cytochromes, and peroxidases. It is also proposed to synthesize a variety of unlabeled heme analogues, heme dimers, iron oxophlorins, and hydrdohemes. The proposed work will provide a basis for understanding heme protein function, and of structural and electronic factors which cause several debilitating diseases such as anemias. The synthetic compounds will also be used for EPR, X-ray, MCD, and other biological studies in collaboration with a number of independent investigators. After reconstruction into appropriate apoproteins, the novel synthetic compounds will be used to produce spectroscopic assignments, and for simplification of spectra and interpretations of a wide variety of heme proteins. Tow fundamental approaches for synthesis of novel compounds will be developed; carbon-13, nitrogen-15, fluorine labeled, and unlabeled prophyrins will be obtained by total synthesis from acyclic precursors. Methods for such approaches have already been worked out, but improvements and discovery of new methodology is planned and anticipated. Some deuterium labeled, carbon-13 labeled porphyrins, protoporphyrin IX analogues, oxophlorins, and sulfhemes, and other chlorin hemes will be approached by manipulation of substituents on existing commercially available porphyrins or from chlorophyll alpha which will be extracted form a commercially available alga. After method discovery and optimization, and in order to fully exploit the synthetic developments which usually take place only on limited scales, we plan to prepare most labeled, unlabeled and structurally modified hemes in batches of approximately 100 mg; these compounds will then be provided to collaborators so that biologically important multinuclear NMR, resonance Raman, EPR, X-ray, MCD, and other properties of the hemes, reconstituted heme proteins, non-iron metal complexes and structural modifications of the porphyrin ligands can be investigated, correlated, rationalized and interpreted.