Development of a high-throughput crystal structure-determination platform for JAK1 using a novel metal-chelator soaking system

Crystals of phosphorylated JAK1 kinase domain were initially generated in complex with nucleotide (ADP) and magnesium. The tightly bound Mg2+-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.

Source: Acta Crystallographica Section F - October 26, 2016 Category: Biochemistry Authors: 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. Tags: Janus kinase kinases crystal soaking ligand exchange structure-based drug design research communications Source Type: research