Download MendeleyDevelopment of a high-throughput crystal structure-determination platform for JAK1 using a novel metal-chelator soaking system.
Caspers, N.L., Han, S., Rajamohan, F., Hoth, L.R., Geoghegan, K.F., Subashi, T.A., Vazquez, M.L., Kaila, N., Cronin, C.N., Johnson, E., Kurumbail, R.G.(2016) Acta Crystallogr F Struct Biol Commun 72: 840-845
- PubMed: 27827355
- DOI: https://doi.org/10.1107/S2053230X16016356
- Primary Citation of Related Structures:
5KHW, 5KHX - PubMed Abstract:
Crystals of phosphorylated JAK1 kinase domain were initially generated in complex with nucleotide (ADP) and magnesium. The tightly bound Mg 2+ -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.
Organizational Affiliation:
Structural Biology, Pfizer Inc., Eastern Point Road, Groton, CT 06340, USA.
