利用变性剂脉冲伴侣蛋白结合构建折叠蛋白的动力学控制变性等温线
Constructing Kinetically Controlled Denaturation Isotherms of Folded Proteins Using Denaturant-Pulse Chaperonin Binding.
作者信息
O'Neil Pierce T, Machen Alexandra J, Thompson Jackie A, Wang Wei, Hoang Quyen Q, Baldwin Michael R, Khar Karen R, Karanicolas John, Fisher Mark T
机构信息
Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas, KS, USA.
Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, IN, USA.
出版信息
Methods Mol Biol. 2019;1873:293-304. doi: 10.1007/978-1-4939-8820-4_19.
Methods to assess the kinetic stability of proteins, particularly those that are aggregation prone, are very useful in establishing ligand induced stabilizing effects. Because aggregation prone proteins are by nature difficult to work with, most solution based methods are compromised by this inherent instability. Here, we describe a label-free method that examines the denaturation of immobilized proteins where the dynamic unfolded protein populations are captured and detected by chaperonin binding.
评估蛋白质动力学稳定性的方法，尤其是那些易于聚集的蛋白质，对于确定配体诱导的稳定作用非常有用。由于易于聚集的蛋白质本质上难以处理，大多数基于溶液的方法都受到这种固有不稳定性的影响。在这里，我们描述了一种无标记方法，该方法通过伴侣蛋白结合捕获并检测固定化蛋白质的变性，从而研究固定化蛋白质的变性情况。
Zotero 插件