可溶性蛋白在类脂立方相中的连续皮秒晶体学。
Serial femtosecond crystallography of soluble proteins in lipidic cubic phase.
机构信息
Center for Applied Structural Discovery at the Biodesign Institute, Department of Chemistry and Biochemistry, Arizona State University , 727 East Tyler Street, Tempe, AZ 85287, USA.
Bridge Institute, Department of Chemistry, University of Southern California , 3430 South Vermont Avenue, MC 3303, Los Angeles, CA 90089, USA.
出版信息
IUCrJ. 2015 Aug 4;2(Pt 5):545-51. doi: 10.1107/S2052252515013160. eCollection 2015 Sep 1.
Serial femtosecond crystallography (SFX) at X-ray free-electron lasers (XFELs) enables high-resolution protein structure determination using micrometre-sized crystals at room temperature with minimal effects from radiation damage. SFX requires a steady supply of microcrystals intersecting the XFEL beam at random orientations. An LCP-SFX method has recently been introduced in which microcrystals of membrane proteins are grown and delivered for SFX data collection inside a gel-like membrane-mimetic matrix, known as lipidic cubic phase (LCP), using a special LCP microextrusion injector. Here, it is demonstrated that LCP can also be used as a suitable carrier medium for microcrystals of soluble proteins, enabling a dramatic reduction in the amount of crystallized protein required for data collection compared with crystals delivered by liquid injectors. High-quality LCP-SFX data sets were collected for two soluble proteins, lysozyme and phycocyanin, using less than 0.1 mg of each protein.
连续飞秒晶体学（SFX）在 X 射线自由电子激光（XFEL）的应用下，使得在室温下使用微米大小的晶体就能实现高分辨率的蛋白质结构测定，同时最小化辐射损伤的影响。SFX 需要源源不断的微晶体以随机取向交叉 XFEL 光束。最近引入了一种 LCP-SFX 方法，其中膜蛋白的微晶体在凝胶状类似膜的基质（称为层状立方相，LCP）中生长并输送，用于 SFX 数据收集，使用特殊的 LCP 微挤出注射器。这里证明，LCP 也可以用作可溶性蛋白质微晶体的合适载体介质，与使用液体注射器输送的晶体相比，大大减少了数据收集所需的结晶蛋白量。使用少于 0.1mg 的每种蛋白质，就成功为两种可溶性蛋白质，溶菌酶和藻蓝蛋白，收集到了高质量的 LCP-SFX 数据集。
Zotero 插件