Structures of ubiquitin insertion mutants support site-specific reflex response to insertions hypothesis.
Structures of ubiquitin insertion mutants support site-specific reflex response to insertions hypothesis.
复制标题
泛素插入突变体的结构支持对插入假说的位点特异性反射反应。
DOI：
10.1016/j.jmb.2006.03.047
复制
发表时间：
2006
影响因子：
5.6
通讯作者：
Robertson,AndrewD
中科院分区：
文献类型：
--
作者：
Ferraro,DebraM;Ferraro,DanielJ;Ramaswamy,S;Robertson,AndrewD
我们以前的结论是，从NMR化学位移判断，插入到泛素的构象的影响似乎主要取决于插入的网站，而不是插入的序列。为了更完整和原子分辨率的理解这些插入如何调节泛素的构象，我们已经解决了四个插入突变体的泛素的晶体结构。泛素残基9和10之间的插入对蛋白质的其余部分的干扰最小，而当插入在残基35和36之间时发生较大的改变。此外，对于给定位点处的每个突变体，响应于插入的改变非常相似。从结构同源蛋白质设计两个插入，每个位点一个。有趣的是，这五到七个氨基酸残基插入的二级结构在新蛋白质中是保守的。总的来说，晶体结构支持先前的结论，这些插入的构象效应主要由插入位点决定，仅由插入序列决定。
We previously concluded that, judging from NMR chemical shifts, the effects of insertions into ubiquitin on its conformation appear to depend primarily on the site of insertion rather than the sequence of the insertion. To obtain a more complete and atomic-resolution understanding of how these insertions modulate the conformation of ubiquitin, we have solved the crystal structures of four insertional mutants of ubiquitin. Insertions between residues 9 and 10 of ubiquitin are minimally perturbing to the remainder of the protein, while larger alterations occur when the insertion is between residues 35 and 36. Further, the alterations in response to insertions are very similar for each mutant at a given site. Two insertions, one at each site, were designed from structurally homologous proteins. Interestingly, the secondary structure within these five to seven amino acid residue insertions is conserved in the new protein. Overall, the crystal structures support the previous conclusion that the conformational effects of these insertions are determined largely by the site of insertion and only secondarily by the sequence of the insert.