Screening of protein kinases by ATP-STD NMR spectroscopy
Screening of protein kinases by ATP-STD NMR spectroscopy
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DOI：
10.1021/ja0425942
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发表时间：
2005-06-08
影响因子：
15
通讯作者：
Wyss, DF
中科院分区：
文献类型：
--
作者：
McCoy, MA;Senior, MM;Wyss, DF
ATP- std NMR利用Mg2+与ATP的结合来调节ATP对蛋白激酶的亲和力，从而可以确定ATP竞争配体的大范围ki 's。用Mn2+取代Mg2+产生顺磁探针（MnATP），由此可以推断非atp竞争配体的接近性。内部标准和参考文献用于减少由于蛋白质或化合物降解而产生的假阳性。使用天然ATP配体赋予活性位点特异性，这是其他配体结合实验无法先验地获得的。
ATP-STD NMR takes advantage of Mg2+binding to ATP to adjust the ATP affinity for protein kinases permitting a wide range ofKi's to be determined for ATP competitive ligands. Substituting Mn2+for Mg2+creates a paramagnetic probe (MnATP) from which the proximity of non-ATP competitive ligands can be inferred. Internal standards and references are used to reduce false positives due to protein or compound degradation. Use of the natural ATP ligand confers active site-specificity that is not available a priori from other ligand binding experiments.