Caspers Nicole L, Han Seungil, Rajamohan Francis, Hoth Lise R, Geoghegan Kieran F, Subashi Timothy A, Vazquez Michael L, Kaila Neelu, Cronin Ciarán N, Johnson Eric, Kurumbail Ravi G
Structural Biology, Pfizer Inc., Eastern Point Road, Groton, CT 06340, USA.
Pharmacokinetics, Dynamics and Metabolism, Pfizer Inc., Eastern Point Road, Groton, CT 06340, USA.
Acta Crystallogr F Struct Biol Commun. 2016 Nov 1;72(Pt 11):840-845. doi: 10.1107/S2053230X16016356. Epub 2016 Oct 27.
Crystals of phosphorylated JAK1 kinase domain were initially generated in complex with nucleotide (ADP) and magnesium. The tightly bound Mg-ADP at the ATP-binding site proved recalcitrant to ligand displacement. Addition of a molar excess of EDTA helped to dislodge the divalent metal ion, promoting the release of ADP and allowing facile exchange with ATP-competitive small-molecule ligands. Many kinases require the presence of a stabilizing ligand in the ATP site for crystallization. This procedure could be useful for developing co-crystallization systems with an exchangeable ligand to enable structure-based drug design of other protein kinases.
磷酸化JAK1激酶结构域的晶体最初是与核苷酸（ADP）和镁形成复合物而产生的。ATP结合位点处紧密结合的Mg-ADP被证明难以被配体置换。加入摩尔过量的EDTA有助于去除二价金属离子，促进ADP的释放，并允许与ATP竞争性小分子配体轻松交换。许多激酶在ATP位点需要存在稳定配体才能结晶。该方法可能有助于开发具有可交换配体的共结晶系统，以实现其他蛋白激酶基于结构的药物设计。
