Biomolecular Structure and Function Group, Institute for Bioscience and Biotechnology Research, National Institute of Standards and Technology and University of Maryland, Rockville, Maryland, USA.
Fischell Department of Bioengineering, University of Maryland, College Park, Maryland, USA.
Proteins. 2020 Sep;88(9):1189-1196. doi: 10.1002/prot.25890. Epub 2020 Mar 27.
ClpS2 is a small protein under development as a probe for selectively recognizing N-terminal amino acids of N-degron peptide fragments. To understand the structural basis of ClpS2 specificity for an N-terminal amino acid, all atom molecular dynamics (MD) simulations were conducted using the sequence of a bench-stable mutant of ClpS2, called PROSS. We predicted that a single amino acid leucine to asparagine substitution would switch the specificity of PROSS ClpS2 to an N-terminal tyrosine over the preferred phenylalanine. Experimental validation of the mutant using a fluorescent yeast-display assay showed an increase in tyrosine binding over phenylalanine, in support of the proposed hypothesis.
ClpS2 是一种正在开发中的小分子蛋白，可用作选择性识别 N 端去稳定肽片段 N-末端氨基酸的探针。为了了解 ClpS2 对 N 端氨基酸特异性的结构基础，使用 ClpS2 的一种稳定突变体 PROSS 的序列进行了全原子分子动力学 (MD) 模拟。我们预测，单个氨基酸亮氨酸到天冬酰胺的取代将使 PROSS ClpS2 的特异性从首选的苯丙氨酸转变为 N 端酪氨酸。使用荧光酵母展示测定法对突变体进行的实验验证显示，酪氨酸结合增加，苯丙氨酸减少，这支持了所提出的假设。
