McCoy Mark A, Senior Mary M, Wyss Daniel F
Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, New Jersey 07033, USA.
J Am Chem Soc. 2005 Jun 8;127(22):7978-9. doi: 10.1021/ja0425942.
ATP-STD NMR takes advantage of Mg2+ binding to ATP to adjust the ATP affinity for protein kinases permitting a wide range of Ki'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.
ATP-STD核磁共振利用镁离子与ATP的结合来调节ATP对蛋白激酶的亲和力，从而能够测定一系列ATP竞争性配体的抑制常数（Ki）。用锰离子取代镁离子可产生顺磁探针（锰-ATP），由此可推断非ATP竞争性配体的接近程度。使用内标和参比物来减少因蛋白质或化合物降解导致的假阳性。使用天然ATP配体赋予了活性位点特异性，这是其他配体结合实验无法预先获得的。
