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Development of a high‐throughput crystal structure‐determination platform for JAK1 using a novel metal‐chelator soaking system
Author(s) -
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.
Publication year - 2016
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x16016356
Subject(s) - ligand (biochemistry) , crystallization , kinase , chemistry , divalent , chelation , metal , crystallography , combinatorial chemistry , stereochemistry , inorganic chemistry , biochemistry , receptor , organic chemistry
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.