Durst Florian G, Ou Horng Der, Löhr Frank, Dötsch Volker, Straub Wesley E
Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance, and Cluster of Excellence Macromolecular Complexes (CEF), University of Frankfurt, Max-von-Laue Strasse 9, 60438 Frankfurt am Main, Germany
J Am Chem Soc. 2008 Nov 12;130(45):14932-3. doi: 10.1021/ja806212j. Epub 2008 Oct 21.
One of the most crucial steps in protein structure determination by nuclear magnetic resonance (NMR) spectroscopy is the preparation of highly concentrated and well behaving protein samples. Here we present a system of modular tags which allows for high level expression, sophisticated purification of full-length protein, and solubility enhancement while keeping the amount of additional resonances low. This system consists of two different expression constructs and utilizes the tight binding of human calmodulin (hCaM) to the calmodulin binding peptide (CBP), which has already been used as a purification tag.
通过核磁共振(NMR)光谱法测定蛋白质结构的最关键步骤之一是制备高浓度且性质良好的蛋白质样品。在此,我们展示了一种模块化标签系统,该系统能够实现高水平表达、对全长蛋白质进行精细纯化以及提高溶解度,同时保持额外共振信号的数量较低。该系统由两种不同的表达构建体组成,并利用人钙调蛋白(hCaM)与钙调蛋白结合肽(CBP)的紧密结合,CBP已被用作纯化标签。