Small-Molecule Phosphine Activation of Protein Function in Zebrafish Embryos with an Expanded Genetic Code.
Small-Molecule Phosphine Activation of Protein Function in Zebrafish Embryos with an Expanded Genetic Code.
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小分子膦激活具有扩展遗传密码的斑马鱼胚胎中的蛋白质功能。
DOI：
10.1007/978-1-0716-3251-2_18
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Deiters,Alexander
中科院分区：
文献类型：
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作者：
Brown,Wes;Rosenblum,Carolyn;Deiters,Alexander
在活的模式生物中对蛋白质功能的条件控制是研究该蛋白质在发育和疾病期间的作用的重要工具。在本章中，我们将详细介绍通过将非经典氨基酸掺入蛋白质活性位点来在斑马鱼胚胎中产生小分子可激活酶的步骤。这种方法可以应用于许多酶类，我们强调与荧光素酶和蛋白酶的时间控制。我们表明，战略布局的非规范氨基酸完全阻断酶的活性，然后迅速恢复后，加入无毒的小分子诱导剂的胚胎水。
Conditional control of protein function in a living model organism is an important tool for studying the effects of that protein during development and disease. In this chapter, we walk through the steps to generate a small-molecule-activatable enzyme in zebrafish embryos through the incorporation of a noncanonical amino acid into the protein active site. This method can be applied to many enzyme classes, which we highlight with temporal control of a luciferase and a protease. We demonstrate that strategic placement of the noncanonical amino acid completely blocks enzyme activity, which is then promptly restored after addition of the nontoxic small molecule inducer to the embryo water.