Chamberlain A K, Fischer K F, Reardon D, Handel T M, Marqusee A S
Department of Molecular and Cell Biology, University of California, Berkeley 94720-3206, USA.
Protein Sci. 1999 Nov;8(11):2251-7. doi: 10.1110/ps.8.11.2251.
Based on results from both equilibrium and kinetic hydrogen exchange studies of Escherichia coli ribonuclease HI (RNase H), a fragment of RNase H (eABCD) was designed. The sequence of eABCD contains less than half of the protein's primary sequence and includes the regions that were shown to be the most protected from hydrogen exchange in all previous studies of RNase H. This core fragment of RNase H encodes a well-ordered protein with native-like properties. When isolated from the full-length monomeric protein, the eABCD fragment forms a stable dimer. However, we show indirectly that the monomeric form of eABCD is folded and has an overall secondary structure similar to the dimeric form.
基于对大肠杆菌核糖核酸酶HI(RNase H)的平衡和动力学氢交换研究结果,设计了RNase H的一个片段(eABCD)。eABCD的序列包含的蛋白质一级序列不到一半,并且包括在之前所有RNase H研究中显示对氢交换最具保护作用的区域。RNase H的这个核心片段编码一种具有天然样性质的有序蛋白质。当从全长单体蛋白中分离出来时,eABCD片段形成稳定的二聚体。然而,我们间接表明,eABCD的单体形式是折叠的,并且具有与二聚体形式相似的整体二级结构。