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

Journal Article · Thu Oct 27 00:00:00 EDT 2016 · Acta Crystallographica. Section F, Structural Biology Communications

DOI:https://doi.org/10.1107/S2053230X16016356· OSTI ID:1405015

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.

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.

• Research Organization: Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
• Sponsoring Organization: INDUSTRY
• OSTI ID: 1405015
• Journal Information: Acta Crystallographica. Section F, Structural Biology Communications, Vol. 72, Issue 11; ISSN 2053-230X
• Publisher: International Union of Crystallography
• Country of Publication: United States
• Language: ENGLISH