A manual nanoscale method for protein crystallization.
A manual nanoscale method for protein crystallization.
复制标题
蛋白质结晶的手动纳米级方法。
DOI：
10.1107/s0907444903011867
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发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Yeh,JoanneI
中科院分区：
文献类型：
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作者：
Yeh,JoanneI
为了克服三维晶体结构测定的主要障碍之一-需要大量纯化材料来生长晶体-结晶方法已经开发出来，只需要总共2-5 μ l的浓缩大分子溶液来筛选100多种条件。这些程序采用圆形载玻片，其中包含25个孔阵列，设计用于在纳升体积范围内的结晶设置。这些“结晶滑块”适合标准结晶托盘的孔。这些纳米级结晶方法已被用于可重复地获得三种蛋白质的良好衍射晶体，其中两种正在积极研究（甘油激酶和NADH过氧化物酶）和一种测试蛋白质（溶菌酶），仅使用40-350µg （0.04-0.35 mg）蛋白质筛选100种条件。这些纳米液结晶方法很容易适用于典型的实验室，不需要机器人或昂贵的设备。
To overcome one of the major hurdles in three-dimensional crystal structure determination – the requirement for large quantities of purified material to grow crystals – crystallization methodologies have been developed that require only a total of 2–5 µl of a concentrated macromolecular solution to screen more than 100 conditions. These procedures employ a circular slide containing an array of 25 wells designed for crystallization setups in the nanolitre volume range. These `crystallization slides' fit into the wells of standard crystallization trays. These nanoscale crystallization approaches have been used to reproducibly obtain well diffracting crystals of three proteins, two that are being actively studied (glycerol kinase and NADH peroxidase) and one test protein (lysozyme), using only 40–350 µg (0.04–0.35 mg) of proteins to screen 100 conditions. These nanolitre crystallization methods are easily adapted for the typical laboratory, without the requirement of robotics or expensive equipment.