SYNTHESES OF LABELED HEMES FOR PROTEIN NMR STUDIES

用于蛋白质 NMR 研究的标记血红素的合成

• 批准号: 3336793
• 负责人: Kevin M Smith
• 金额: $ 15.68万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-04-01 至 1991-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3336793
• 关键词: Raman spectrometry; X ray crystallography; catalase; chemical structure function; chemical synthesis; cytochromes; deuterium; electron spin resonance spectroscopy; fluorine; heme; hemoglobin; hemoprotein; hemoprotein structure; high performance liquid chromatography; ligands; metalloproteins; myoglobin; nuclear magnetic resonance spectroscopy; peroxidases; porphyrins; protein structure function; radiotracer; tritium

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.

建议开发和利用方法， 合成天然血红素的预定区域选择性标记 具有氘、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，以获得一个 了解血红素载脂蛋白相互作用和结构/功能 在多种生物学上重要的血红素蛋白中的关系， 血红蛋白、肌红蛋白、细胞色素和过氧化物酶。 还拟议 合成各种未标记的血红素类似物、血红素二聚体、铁 氧代叶绿素和氢血红素。 拟议的工作将为以下方面提供基础： 了解血红素蛋白的功能，以及结构和电子 导致贫血等多种衰弱性疾病的因素。 的 合成化合物也将用于EPR，X射线，MCD和其他 生物学研究与一些独立的 investigators. 在重组成适当的脱辅基蛋白后， 新的合成化合物将用于生产光谱 分配，并简化光谱和解释的一个 种类繁多的血红素蛋白。 两种基本的综合方法 将开发多种新型化合物;碳-13、氮-15、氟 标记的和未标记的野百合素将通过全合成从 无环前体 这种方法已经被研究出来了 但计划改进和发现新的方法， 预期。 一些氘标记的，碳-13标记的卟啉， 原卟啉IX类似物、氧代卟啉和硫代血红素以及其他二氢卟酚 血红素将通过操纵现有的取代基来处理。 可商购的卟啉或来自叶绿素α， 提取自市售可得的乙醇。 方法发现后， 优化，为了充分利用合成的发展， 这通常只发生在有限的规模，我们计划准备最 标记的、未标记的和结构修饰的血红素， 大约100 mg;然后将这些化合物提供给 合作者，使生物重要的多核核磁共振，共振 血红素的拉曼、EPR、X射线、MCD和其他性质， 血红素蛋白、非铁金属络合物和血红素蛋白的结构修饰 卟啉配体可以被研究、关联、合理化并 翻译。