Development of a high-throughput crystal structure-determination platform for JAK1 using a novel metal-chelator soaking system.
Acta Crystallogr F Struct Biol Commun
; 72(Pt 11): 840-845, 2016 11 01.
Article
em En
| MEDLINE
| ID: mdl-27827355
ABSTRACT
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.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Temas:
Geral
Base de dados:
MEDLINE
Assunto principal:
Difosfato de Adenosina
/
Trifosfato de Adenosina
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Ácido Edético
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Cristalização
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Janus Quinase 1
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Magnésio
Limite:
Animals
/
Humans
Idioma:
En
Revista:
Acta Crystallogr F Struct Biol Commun
Ano de publicação:
2016
Tipo de documento:
Article
País de afiliação:
Estados Unidos