Shah Premal S, Hom Geoffrey K, Ross Scott A, Lassila Jonathan Kyle, Crowhurst Karin A, Mayo Stephen L
Biochemistry and Molecular Biophysics Option, MC 114-96, California Institute of Technology, Pasadena, CA 91125, USA.
J Mol Biol. 2007 Sep 7;372(1):1-6. doi: 10.1016/j.jmb.2007.06.032. Epub 2007 Jun 16.
Computational protein design procedures were applied to the redesign of the entire sequence of a 51 amino acid residue protein, Drosophila melanogaster engrailed homeodomain. Various sequence optimization algorithms were compared and two resulting designed sequences were experimentally evaluated. The two sequences differ by 11 mutations and share 22% and 24% sequence identity with the wild-type protein. Both computationally designed proteins were considerably more stable than the naturally occurring protein, with midpoints of thermal denaturation greater than 99 degrees C. The solution structure was determined for one of the two sequences using multidimensional heteronuclear NMR spectroscopy, and the structure was found to closely match the original design template scaffold.
计算蛋白质设计程序被应用于对一个由51个氨基酸残基组成的蛋白质——果蝇成对控制基因同源异型结构域的全序列进行重新设计。比较了各种序列优化算法，并对由此产生的两个设计序列进行了实验评估。这两个序列有11个突变差异，与野生型蛋白质的序列同一性分别为22%和24%。两种通过计算设计的蛋白质都比天然存在的蛋白质稳定得多，热变性中点温度高于99摄氏度。使用多维异核核磁共振光谱法确定了两个序列之一的溶液结构，发现该结构与原始设计模板支架紧密匹配。
