SELENO METHIONINE INCORPORATION IN RECOMBINANT PROTEINS FOR XRAY CRYSTALLOGRAPHY

我们的研究主要集中在其结构和功能上。 载脂蛋白(Apo)E和其他与心脏和心脏相关的蛋白质 阿尔茨海默病。我们的主要结构工具是x射线。 结晶学。为了提高我们解决新问题的效率 结构，并避免使用多个同构 为了解决相位问题，我们开发了重组 硒蛋氨酸可以利用的蛋白质表达系统 甲硫氨酸的替代品。有了这种替代，我们可以分阶段 利用多重反常技术直接在单晶上实现 分散度(MAD)。我们建议使用加州大学旧金山分校的质谱仪 确认将硒蛋氨酸加入我们的 在进行结晶和数据之前的重组蛋白质 收集，这将在同步加速器的MAD站完成 设施。

Our research focuses on the structure and function of apolipoprotein (apo) E and other proteins involved in heart and Alzheimer's diseases. Our main structural tool is x-ray crystallography. In order to increase our efficiency in solving new structures and avoiding the needto use multiple isomorphous replacement to solve the phase problem, we have developed recombinant protein expression systems with which seleno-methionine can be substituted for methionine. With this substitution we can phase directly on a single crystal using the technique of multiple anomalous dispersion (MAD). We propose to use the UCSF Mass Spectrometry Facility to confirm the incorporation of seleno-methionine into our recombinant proteins before proceeding with crystallization and data collection, which will be done at a MAD station at a synchrotron facility.