FLEXIQinase, a mass spectrometry-based assay, to unveil multikinase mechanisms.
Nat Methods
; 9(5): 504-8, 2012 Apr 08.
Article
em En
| MEDLINE
| ID: mdl-22484849
ABSTRACT
We introduce a mass spectrometry-based method that provides residue-resolved quantitative information about protein phosphorylation. In this assay we combined our full-length expressed stable isotope-labeled protein for quantification strategy (FLEXIQuant) with a traditional kinase assay to determine the mechanisms of multikinase substrate phosphorylation such as priming-dependent kinase activities. The assay monitors the decrease in signal intensity of the substrate peptides and the concomitant increase in the (n × 80 Da)-shifted phosphorylated peptide. We analyzed the c-Jun N-terminal kinase (JNK)-dependent glycogen synthase kinase 3ß (GSK3ß) activity on doublecortin (DCX) revealing mechanistic details about the role of phosphorylation cross-talk in GSK3ß activity and permitting an advanced model for GSK3ß-mediated signaling.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Neuropeptídeos
/
Quinase 3 da Glicogênio Sintase
/
Proteínas Quinases JNK Ativadas por Mitógeno
/
Espectrometria de Massas em Tandem
/
Proteínas Associadas aos Microtúbulos
Idioma:
En
Revista:
Nat Methods
Assunto da revista:
TECNICAS E PROCEDIMENTOS DE LABORATORIO
Ano de publicação:
2012
Tipo de documento:
Article
País de afiliação:
Estados Unidos