A simple way to measure protein refolding rates in water.
A simple way to measure protein refolding rates in water.
复制标题
测量水中蛋白质重折叠率的简单方法。
DOI：
10.1006/jmbi.2001.5039
复制
发表时间：
2001
影响因子：
5.6
通讯作者：
M. Oliveberg
中科院分区：
文献类型：
--
作者：
D. Otzen;M. Oliveberg
Refolding of proteins is traditionally carried out either by diluting the denaturant-unfolded protein into buffer (GdmCl-jump) or by mixing the acid-denatured protein with strong buffer (pH-jump). The first method does not allow direct measurement of folding rates in water since the GdmCl cannot be infinitely diluted, and the second method suffers from the limitation that many proteins cannot be pH-denatured. Further, some proteins do not refold reversibly from low pH where they get trapped as aggregation prone intermediates. Here, we present an alternative approach for direct measurement of refolding rates in water, which does not rely on extrapolation. The protein is denatured in SDS, and is then mixed with alpha-cyclodextrin, which rapidly strips SDS molecules from the protein, leaving the naked unfolded protein to refold.
