XRAY DIFFRACTION OF CLATHRIN HUB CRYSTALS

点击翻译按钮获取中文摘要

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.