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: 3336783
• 负责人: Kevin M Smith
• 金额: $ 15.89万
• 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/3336783
• 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，以允许 天然血液的合成区域选择性标记为预定的 氘，碳-13，tritium，氮15和氟的位置， 这些将用作NMR和共振拉曼探针，以获得 理解血红素的载蛋白相互作用和结构/功能 各种生物学上重要的血红素蛋白的关系，例如 血红蛋白，肌球蛋白，细胞色素和过氧化物酶。 也提出了 合成各种未标记的血红素类似物，血红素二聚体，铁 氧氯蛋白和水力局。 拟议的工作将为 了解血红素蛋白功能以及结构和电子 引起多种令人衰弱的疾病（例如贫血）的因素。 这 合成化合物也将用于EPR，X射线，MCD和其他 生物学研究与许多独立 调查人员。 重建为适当的载蛋白后， 新型合成化合物将用于产生光谱 作业，并简化光谱和解释 多种血红素蛋白。 合成的基本方法 将开发新的化合物；碳13，氮15，氟 标记的，未标记的预知蛋白将通过总合成而获得 无环前体。 这种方法的方法已经有效 出去，但是计划了新方法的改进和发现，并且 预期。 一些标记为碳13标记卟啉的氘， 原源性IX类似物，氧氯蛋白和硫酸盐和其他氯蛋白 在现有 商业可用的卟啉或叶绿素α将是 提取的形式是商业上可用的藻类。 方法发现和 优化，并为了充分利用合成发展 通常仅在有限的尺度上进行，我们计划准备大多数 标记，未标记和结构修饰的下摆 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NMR共振 拉曼，EPR，X射线，MCD和HEMES的其他特性，重组 血红素蛋白，非铁金属复合物和结构修饰 可以研究，相关，合理化的卟啉配体 解释。