SYNTHESES OF LABELED HEMES FOR PROTEIN NMR STUDIES

建议开发和开发方法学，以使 区域选择性标记天然血红素的合成 含氢、碳-13、氚、氮-15和氟的位置， 这些将被用作核磁共振和共振拉曼探针，以获得 关于血红素载脂蛋白相互作用和结构/功能的认识 各种生物重要的血红素蛋白之间的关系，如 血红蛋白、肌红蛋白、细胞色素和过氧化物酶。它还被提议 合成各种未标记的血红素类似物、血红素二聚体、铁 氧化苦参碱和氢化血红素。拟议的工作将为 了解血红素蛋白的功能，以及结构和电子结构 导致几种衰弱疾病的因素，如贫血。这个 合成化合物还将用于EPR、X射线、MCD和其他 与多个独立的生物研究机构合作 调查人员。在重建成适当的脱辅基蛋白后， 新型合成化合物将用于生产光谱 指认，并简化光谱和解释一个 种类繁多的血红素蛋白质。合成的两种基本方法 将开发一系列新的化合物；碳-13，氮-15，氟 标记的和未标记的卟啉将通过全合成得到 非环前体物。用于这种方法的方法已经被研究出来了 ，但是改进和发现新的方法是计划的，并且 已经预料到了。一些氢标记的，碳-13标记的卟啉， 原卟啉IX类似物、氧杂卟啉、亚硫胺和其他氯 通过操纵现有的取代基来接近亚铁血红素 商业上可获得的卟啉或来自叶绿素α的将是 从一种商业上可用的藻类中提取。在方法发现和 优化，为了充分利用综合开发 通常只在有限的规模上进行，我们计划准备大多数 批次标记、未标记和结构修饰的大麻 大约100毫克；然后这些化合物将被提供给 合作者使具有重要生物意义的多核核磁共振、共振 重组的血红素的拉曼、EPR、X-射线、MCD等性质 血红素蛋白、非铁金属络合物及其结构修饰 可以对卟啉配体进行研究、关联、合理化和 翻译过来了。

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.