XRAY DIFFRACTION OF CLATHRIN HUB CRYSTALS

网格蛋白中心晶体的 X 射线衍射

• 批准号: 6658685
• 负责人: ROBERT J FLETTERICK
• 金额: $ 14.32万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-01 至 2003-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6658685
• 关键词: bioimaging /biomedical imaging; biomaterials; biomedical resource; endocrine gland /system; proteins; radiography; structural biology

Clathrin-coated vesticles (CCV) mediate endocytosis of receptor-bound nutrients, hormones and proteins destined for degradation, and regulate physiological levels of these ligands. CCV mediates sorting processes in the trans-Golgi network required for biogenesis of lysosomes and secretary granules. We have expressed, purified and crystallized the clathrin hub/LCb complex which contains residues 1074-1675 of the heavy chain and neuronal Lcb light chain. Studies demonstrated that the hub/LCb complex contains all information needed for assembly. Elucidation of the structure will reveal the molecular mechanism for assembly process. The crystals we have are too small to get high enough resolution data on a rotating anode generator for a structure determination. We would like to get diffraction to 3 E resolution or beyond at a synchrotron radiation and, collect a complete native data set. We have determined the conditions to freeze the crystals with minor effect on the mosaicity of the diffraction pattern.

网格蛋白包被的囊泡（CCV）介导 受体结合的营养素、激素和蛋白质， 降解，并调节这些配体的生理水平。 CCV 介导的排序过程中所需的trans-Golgi网络 溶酶体和分泌颗粒的生物发生。 我们已经表示， 纯化并结晶网格蛋白中心/LCb复合物，其含有 重链和神经元Lcb轻链的残基1074-1675。 研究表明，枢纽/LCb复合体包含所有信息， 需要组装。 结构解析将揭示 组装过程的分子机制。 我们拥有的水晶 太小而无法在旋转阳极上获得足够高的分辨率数据 用于结构确定的生成器。 我们希望 在同步辐射下衍射到3E分辨率或更高 并收集完整的原生数据集。 吾等已厘定 冷冻晶体的条件对镶嵌性的影响较小 的衍射图案。