Covalent capture of kinase-specific phosphopeptides reveals Cdk1-cyclin B substrates.
Proc Natl Acad Sci U S A
; 105(5): 1442-7, 2008 Feb 05.
Article
en En
| MEDLINE
| ID: mdl-18234856
ABSTRACT
We describe a method for rapid identification of protein kinase substrates. Cdk1 was engineered to accept an ATP analog that allows it to uniquely label its substrates with a bio-orthogonal phosphate analog tag. A highly specific, covalent capture-and-release methodology was developed for rapid purification of tagged peptides derived from labeled substrate proteins. Application of this approach to the discovery of Cdk1-cyclin B substrates yielded identification of >70 substrates and phosphorylation sites. Many of these sites are known to be phosphorylated in vivo, but most of the proteins have not been characterized as Cdk1-cyclin B substrates. This approach has the potential to expand our understanding of kinase-substrate connections in signaling networks.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Fosfopéptidos
/
Espectrometría de Masas
/
Proteína Quinasa CDC2
/
Ciclina B
Tipo de estudio:
Evaluation_studies
Límite:
Humans
Idioma:
En
Revista:
Proc Natl Acad Sci U S A
Año:
2008
Tipo del documento:
Article
País de afiliación:
Estados Unidos