磷酸化形式的CheY的稳定类似物的合成：膦酰基-CheY。
Synthesis of a Stable Analog of the Phosphorylated Form of CheY: Phosphono-CheY.
作者信息
Lookadoo Daniel B, Beyersdorf Matthew S, Halkides Christopher J
机构信息
Department of Chemistry and Biochemistry, The University of North Carolina Wilmington, Wilmington, NC, USA.
出版信息
Methods Mol Biol. 2018;1729:337-343. doi: 10.1007/978-1-4939-7577-8_26.
CheY is a response regulator of bacterial chemotaxis that is activated by phosphorylation of a conserved aspartate residue. However, studies of CheY-phosphate have proven challenging due to rapid hydrolysis of the aspartyl-phosphate in vitro. To combat this issue, we have designed a stable analog suitable for structural and functional studies. Herein, we describe a method for the chemical modification of Thermotoga maritima CheY to produce a phospho-analog designated as phosphono-CheY. Our modification produces a stable analog in the constitutively active form that enables the study of signal transfer to the downstream target.
CheY是细菌趋化作用的一种应答调节因子，通过保守天冬氨酸残基的磷酸化被激活。然而，由于体外天冬氨酰磷酸的快速水解，对磷酸化CheY的研究已被证明具有挑战性。为了解决这个问题，我们设计了一种适用于结构和功能研究的稳定类似物。在此，我们描述了一种对海栖热袍菌CheY进行化学修饰以产生一种称为膦酰基-CheY的磷酸类似物的方法。我们的修饰产生了一种组成型活性形式的稳定类似物，能够研究信号向下游靶点的传递。
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