Methods for discovering catalytic activities for pseudokinases.
Methods for discovering catalytic activities for pseudokinases.
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DOI：
10.1016/bs.mie.2022.03.047
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发表时间：
2022
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假酶类似于活性酶，但缺乏被认为是活性所需的关键催化残基。许多假酶在常规酶测定中似乎是无活性的。然而，对它们明显缺乏活性的另一种解释是，假酶被检测出错误的反应。我们已经发现了几个新的蛋白激酶样家族，这些家族揭示了三磷酸腺苷（ATP）和活性位点残基迁移的不同结合方向如何从共同的激酶支架产生新的反应。这些结果暴露了蛋白激酶折叠的催化多功能性，并建议应分析非典型激酶和假激酶的替代转移酶活性。在这一章中，我们讨论了一个生物信息学识别的酶超家族的分歧或非典型成员的一般方法，然后提出了一个实验方法来表征其催化活性。
Pseudoenzymes resemble active enzymes, but lack key catalytic residues believed to be required for activity. Many pseudoenzymes appear to be inactive in conventional enzyme assays. However, an alternative explanation for their apparent lack of activity is that pseudoenzymes are being assayed for the wrong reaction. We have discovered several new protein kinase-like families which have revealed how different binding orientations of adenosine triphosphate (ATP) and active site residue migration can generate a novel reaction from a common kinase scaffold. These results have exposed the catalytic versatility of the protein kinase fold and suggest that atypical kinases and pseudokinases should be analyzed for alternative transferase activities. In this chapter, we discuss a general approach for bioinformatically identifying divergent or atypical members of an enzyme superfamily, then present an experimental approach to characterize their catalytic activity.