Optimization of protein buffer cocktails using Thermofluor
Optimization of protein buffer cocktails using Thermofluor
复制标题
DOI：
10.1107/s1744309112051858
复制
发表时间：
2013-02-01
影响因子：
0.9
通讯作者：
Weiss, Manfred S.
中科院分区：
文献类型：
--
作者：
Reinhard, Linda;Mayerhofer, Hubert;Weiss, Manfred S.
蛋白质样品的稳定性和均匀性受到蛋白质所在缓冲液组成的强烈影响。鉴定增加蛋白质样品的稳定性和可能的构象均一性的缓冲液组合物的一种快速且简单的方法是基于荧光的热位移测定（Thermofluor）。在这里，一个新的96条件屏幕的Thermofluor实验，它包括缓冲和添加剂部分。缓冲液筛选包括23种不同的缓冲液，添加剂筛选包括小分子添加剂，如盐和核苷酸类似物。利用增加蛋白质样品热稳定性的小分子组分通常导致更高质量和数量的蛋白质制备物，并且最终也增加了蛋白质结晶的机会。
The stability and homogeneity of a protein sample is strongly influenced by the composition of the buffer that the protein is in. A quick and easy approach to identify a buffer composition which increases the stability and possibly the conformational homogeneity of a protein sample is the fluorescence-based thermal-shift assay (Thermofluor). Here, a novel 96-condition screen for Thermofluor experiments is presented which consists of buffer and additive parts. The buffer screen comprises 23 different buffers and the additive screen includes small-molecule additives such as salts and nucleotide analogues. The utilization of small-molecule components which increase the thermal stability of a protein sample frequently results in a protein preparation of higher quality and quantity and ultimately also increases the chances of the protein crystallizing.