Hydroxyethyl cellulose matrix applied to serial crystallography.
Hydroxyethyl cellulose matrix applied to serial crystallography.
复制标题
DOI：
10.1038/s41598-017-00761-0
复制
发表时间：
2017-04-06
影响因子：
4.6
通讯作者：
Iwata S
中科院分区：
文献类型：
--
作者：
Sugahara M;Nakane T;Masuda T;Suzuki M;Inoue S;Song C;Tanaka R;Nakatsu T;Mizohata E;Yumoto F;Tono K;Joti Y;Kameshima T;Hatsui T;Yabashi M;Nureki O;Numata K;Nango E;Iwata S
连续飞秒结晶学(SFX)允许在室温下以最小的辐射损伤来确定蛋白质的结构。高粘度的基质用作晶体载体，用于以低流速加载连续样品，从而能够确定结构，同时需要消耗不到1毫克的样品。然而，对于各种各样的蛋白质样品，可靠和通用的载体基质仍然是难以捉摸的。在这里，我们介绍了羟乙基纤维素基质载体，以确定三种蛋白质的结构。利用Pr原子的反常信号，利用3,000幅衍射图，从单波长反常衍射(SAD)中确定了蛋白酶K的从头结构。
Serial femtosecond crystallography (SFX) allows structures of proteins to be determined at room temperature with minimal radiation damage. A highly viscous matrix acts as a crystal carrier for serial sample loading at a low flow rate that enables the determination of the structure, while requiring consumption of less than 1 mg of the sample. However, a reliable and versatile carrier matrix for a wide variety of protein samples is still elusive. Here we introduce a hydroxyethyl cellulose-matrix carrier, to determine the structure of three proteins. The de novo structure determination of proteinase K from single-wavelength anomalous diffraction (SAD) by utilizing the anomalous signal of the praseodymium atom was demonstrated using 3,000 diffraction images.