Sauer David B, Karpowich Nathan K, Song Jin Mei, Wang Da-Neng
Department of Cell Biology, The Helen L. and Martin S. Kimmel Center for Biology and Medicine, Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, New York.
Department of Cell Biology, The Helen L. and Martin S. Kimmel Center for Biology and Medicine, Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, New York.
Biophys J. 2015 Oct 6;109(7):1420-8. doi: 10.1016/j.bpj.2015.07.026.
Ex vivo stability is a valuable protein characteristic but is laborious to improve experimentally. In addition to biopharmaceutical and industrial applications, stable protein is important for biochemical and structural studies. Taking advantage of the large number of available genomic sequences and growth temperature data, we present two bioinformatic methods to identify a limited set of amino acids or positions that likely underlie thermostability. Because these methods allow thousands of homologs to be examined in silico, they have the advantage of providing both speed and statistical power. Using these methods, we introduced, via mutation, amino acids from thermoadapted homologs into an exemplar mesophilic membrane protein, and demonstrated significantly increased thermostability while preserving protein activity.
体外稳定性是一种有价值的蛋白质特性，但通过实验来改善却很费力。除了生物制药和工业应用外，稳定的蛋白质对于生化和结构研究也很重要。利用大量可用的基因组序列和生长温度数据，我们提出了两种生物信息学方法，以识别一组有限的可能是热稳定性基础的氨基酸或位置。由于这些方法允许在计算机上检查数千个同源物，它们具有提供速度和统计能力的优势。使用这些方法，我们通过突变将来自热适应同源物的氨基酸引入到一个典型的嗜温膜蛋白中，并证明在保留蛋白质活性的同时显著提高了热稳定性。
