Synthesis of Macfarlandin E and Investigation of Localized Golgi Vesiculation

描述(申请人提供)：高尔基体分裂成小泡是细胞分裂所必需的。已经观察到，用结构上有兴趣的海洋天然产物Macfarlandin E处理会导致高尔基体不可逆的碎裂，而不会导致所产生的碎片的非局部化。这项研究的第一个目标是化学合成macfarlandin E，这本身就是对一类不寻常的海洋海绵代谢物的重要贡献。所提出的化学的关键转变是设置两个相邻的立体中心的共轭加成(一个是四元)和分子内捕获氧卡宾离子以建立独特的2，7-二氧双环[3.2.1]Octan-3-酮-在几个海绵天然产物中发现的支架。此外，该合成允许制备包含生物素亲和标签和香豆素部分的类似物，用于研究和分离负责囊泡形成和定位的高尔基体蛋白。本研究的第二个目标是确定Macfarlandin E的生物学靶点，并确定其在调节高尔基体囊泡形成和定位中的作用。了解负责高尔基体行为的成分不仅是对高尔基体生物学的重要贡献，也是更全面地了解导致有丝分裂的细胞过程的第一步。这项拟议的研究涉及海洋天然产物Macfarlandin E及其类似物的化学合成。初步工作表明，macfarlandin E导致哺乳动物细胞中高尔基体不寻常而有趣的碎裂，而不是片段的非定位化。这些化合物将被用作化学试剂，用于研究高尔基体碎裂的性质。

DESCRIPTION (provided by applicant): Fragmentation of the Golgi into small vesicles is a requirement for cellular division. It has been observed that treatment with a structurally interesting marine natural product, macfarlandin E, causes irreversible fragmentation of the Golgi, without causing delocalization of the resulting fragments. The first goal of this research is a chemical synthesis of macfarlandin E, which in itself would be an important contribution to an unusual class of marine spongian metabolites. The key transformations of the proposed chemistry are a conjugate addition to set two adjacent stereocenters (one being quaternary) and a intramolecular trapping of a oxocarbenium ion to build a unique 2,7-dioxabicyclo[3.2.1]octan-3-one scaffold found in several spongian natural products. Additionally, the synthesis allows for preparation of analogs containing biotin affinity tags and coumarin moieties for investigation and isolation of the Golgi proteins responsible for vesiculation and localization. The second goal of the research is to identify the biological target of macfarlandin E and determine its role in the regulation of vesiculation and localization of the Golgi. Understanding the components responsible for Golgi behavior would not only be an important contribution to Golgi biology, but it is the first step toward a more complete understanding of the cellular processes leading up to mitosis. The proposed research involves the chemical synthesis of a marine natural product, macfarlandin E, and its analogs. Preliminary work indicates macfarlandin E causes an unusual and interesting fragmentation of the Golgi aparatus in mammalian cells without delocalization of the fragments. These compounds will be used as chemical agents for the investigation of the nature of Golgi fragmentation.