The Helen L. and Martin S. Kimmel Center for Biology and Medicine at the Skirball Institute of Biomolecular Medicine, Department of Cell Biology, New York University School of Medicine, 540 First Avenue, New York, NY 10016, USA.
Methods. 2011 Dec;55(4):324-9. doi: 10.1016/j.ymeth.2011.07.008. Epub 2011 Aug 5.
Biochemical and biophysical analysis on integral membrane proteins often requires monodisperse and stable protein samples. Here we describe a method to characterize protein thermostability by measuring its melting temperature in detergent using analytical size-exclusion chromatography. This quantitative method can be used to screen for compounds and conditions that stabilize the protein. With this technique we were able to assess and improve the thermostability of several membrane proteins. These conditions were in turn used to assist purification, to identify protein ligand and to improve crystal quality.
生化和生物物理分析整体膜蛋白通常需要单分散和稳定的蛋白质样品。在这里，我们描述了一种通过使用分析尺寸排阻色谱法在去污剂中测量其熔点来测定蛋白质热稳定性的方法。这种定量方法可用于筛选稳定蛋白质的化合物和条件。通过该技术，我们能够评估和提高几种膜蛋白的热稳定性。这些条件反过来又被用于辅助蛋白质的纯化、鉴定蛋白质配体和改善晶体质量。
