Rasia Rodolfo M, Noirclerc-Savoye Marjolaine, Bologna Nicolás G, Gallet Benoit, Plevin Michael J, Blanchard Laurence, Palatnik Javier F, Brutscher Bernhard, Vernet Thierry, Boisbouvier Jérôme
Institut de Biologie Structurale Jean-Pierre Ebel, Centre National de la Recherche Scientifique, Université Joseph-Fourier, Grenoble, France.
Protein Sci. 2009 Feb;18(2):434-9. doi: 10.1002/pro.46.
A major challenge in structural biology remains the identification of protein constructs amenable to structural characterization. Here, we present a simple method for parallel expression, labeling, and purification of protein constructs (up to 80 kDa) combined with rapid evaluation by NMR spectroscopy. Our approach, which is equally applicable for manual or automated implementation, offers an efficient way to identify and optimize protein constructs for NMR or X-ray crystallographic investigations.
结构生物学中的一个主要挑战仍然是鉴定适合进行结构表征的蛋白质构建体。在此,我们提出了一种简单的方法,用于蛋白质构建体(高达80 kDa)的平行表达、标记和纯化,并结合核磁共振光谱进行快速评估。我们的方法同样适用于手动或自动操作,为鉴定和优化用于核磁共振或X射线晶体学研究的蛋白质构建体提供了一种有效的途径。