HotSpot Wizard is a web server used for the automated identification of hotspots in semi-rational protein design to give improved protein stability, catalytic activity, substrate specificity and enantioselectivity. Since there are three orders of magnitude fewer protein structures than sequences in bioinformatic databases, the major limitation to the usability of previous versions was the requirement for the protein structure to be a compulsory input for the calculation. HotSpot Wizard 3.0 now accepts the protein sequence as input data. The protein structure for the query sequence is obtained either from eight repositories of homology models or is modeled using Modeller and I-Tasser. The quality of the models is then evaluated using three quality assessment tools—WHAT_CHECK, PROCHECK and MolProbity. During follow-up analyses, the system automatically warns the users whenever they attempt to redesign poorly predicted parts of their homology models. The second main limitation of HotSpot Wizard’s predictions is that it identifies suitable positions for mutagenesis, but does not provide any reliable advice on particular substitutions. A new module for the estimation of thermodynamic stabilities using the Rosetta and FoldX suites has been introduced which prevents destabilizing mutations among pre-selected variants entering experimental testing. HotSpot Wizard is freely available at http://loschmidt.chemi.muni.cz/hotspotwizard.

中文翻译：

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HotSpot Wizard 3.0：用于基于序列输入信息自动设计突变和智能库的Web服务器

HotSpot Wizard是一种Web服务器，用于在半理性蛋白质设计中自动识别热点，以提高蛋白质的稳定性，催化活性，底物特异性和对映选择性。由于生物信息数据库中的蛋白质结构比序列少三个数量级，因此以前版本的可用性的主要限制是要求蛋白质结构必须作为计算的强制输入。HotSpot Wizard 3.0现在接受蛋白质序列作为输入数据。查询序列的蛋白质结构可以从八个同源性模型库中获得，也可以使用Modeller和I-Tasser进行建模。然后使用三个质量评估工具-WHAT_CHECK，PROCHECK和MolProbity评估模型的质量。在后续分析中，每当用户尝试重新设计其同源性模型预测不良的部分时，系统都会自动向用户发出警告。HotSpot向导的预测的第二个主要限制是它可以确定诱变的合适位置，但不能提供有关特定替代的任何可靠建议。引入了使用Rosetta和FoldX套件估算热力学稳定性的新模块，该模块可防止进入实验测试的预选变体之间的不稳定突变。可以从http://loschmidt.chemi.muni.cz/hotspotwizard免费获得HotSpot向导。但未提供有关特定替代品的任何可靠建议。引入了使用Rosetta和FoldX套件估算热力学稳定性的新模块，该模块可防止进入实验测试的预选变体之间的不稳定突变。可以从http://loschmidt.chemi.muni.cz/hotspotwizard免费获得HotSpot向导。但未提供有关特定替代品的任何可靠建议。引入了使用Rosetta和FoldX套件估算热力学稳定性的新模块，该模块可防止进入实验测试的预选变体之间的不稳定突变。可以从http://loschmidt.chemi.muni.cz/hotspotwizard免费获得HotSpot向导。