精准预测蛋白质稳定性的强大工具

HotSpot Wizard 3是一个网络服务器用于自动设计突变和智能库用于蛋白质功能和稳定性的工程以及蛋白质结构的注释。诱变热点的识别基于从几个生物信息学数据库和计算工具中获得的结构、功能和进化信息的整合。HotSpot Wizard 3 is a web server for an automated design of mutations and smart libraries for the engineering of protein function and stability and the annotation of protein structures.The identification of mutagenesis hot spots is based on the integration of structural, functional and evolutionary information obtained from several bioinformatics databases and computational tools.用户可以探索由四种不同的蛋白质工程策略识别的热点i由位于催化口袋和/或通道中的高度可变残基表示的功能热点ii由柔性残基表示的稳定性热点结构灵活性方法iii由序列同源物中经常被相同残基占据的位置表示的稳定性热点序列共识方法以及iv由调节酶活性和选择性的成对共同进化残基表示的相关热点。Users can explore hot spots identified by four different protein engineering strategies:(i) functional hot spotsrepresented by highly mutable residues located in the catalytic pocket and/or access tunnels,(ii) stability hot spots (structural flexibility approach)represented by flexible residues,(iii) stability hot spots (sequence consensus approach)represented by positions which are frequently occupied by the same residue in sequence homologs, and(iv) correlated hot spotsrepresented by pairs of coevolving residues that modulate enzyme activity and selectivity.对于每种策略HotSpot Wizard列出了根据其预测的诱变适用性排序的残基以及相关的分析特定信息并提供了过滤器使用户可以将识别的热点集减少到最有希望的候选者或根据其标准选择热点。每个位置的详细注释还包括序列同源物中的氨基酸频率和预测的耐受氨基酸可用于选择单个热点的合适替代方案并直接通过HotSpot Wizard界面设计适当的简并密码子。热点以及任何其他残留物都可以在结构中与识别的口袋和隧道一起可视化以探索它们的结构背景。