通过跨尿素梯度等电聚焦和二维等电聚焦研究蛋白质折叠与聚集。

Protein folding and aggregation studied by isoelectric focusing across a urea gradient and isoelectric focusing in two dimensions.

作者信息

van den Oetelaar P J, de Man B M, Hoenders H J

机构信息

Department of Biochemistry, University of Nijmegen, The Netherlands.

出版信息

Biochim Biophys Acta. 1989 Mar 16;995(1):82-90. doi: 10.1016/0167-4838(89)90237-9.

DOI:10.1016/0167-4838(89)90237-9

PMID:2923918

Abstract

Isoelectric focusing across a concentration gradient of urea was used to study the folding-unfolding and association-dissociation processes of proteins. Myoglobulin, albumin, RNase, papain, beta L- and alpha-crystallin were analyzed with this technique, and examples are given of visualized dissociation steps and of equilibrium-unfolding intermediates. Furthermore, a two-dimensional isoelectric focusing technique is presented that is useful to deduce whether a transition of a protein aggregate observed upon urea-gradient isoelectric focusing must be attributed to a change in the protein's tertiary or quaternary structure.

摘要

利用在尿素浓度梯度上进行等电聚焦来研究蛋白质的折叠-去折叠以及缔合-解离过程。用该技术分析了肌红蛋白、白蛋白、核糖核酸酶、木瓜蛋白酶、β-L-晶体蛋白和α-晶体蛋白，并给出了可视化的解离步骤和平衡去折叠中间体的实例。此外，还介绍了一种二维等电聚焦技术，该技术有助于推断在尿素梯度等电聚焦过程中观察到的蛋白质聚集体的转变是否一定归因于蛋白质三级或四级结构的变化。