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Isotipos de proteína quinasa C controlados por la fosfoinositida 3-quinasa a través de la proteína quinasa PDK1
J A Le Good1, W H Ziegler, D B Parekh
1Protein Phosphorylation Laboratory, Imperial Cancer Research Fund, 44 Lincoln's Inn Fields, London WC2A 3PX, UK.
Science (New York, N.Y.)
|September 25, 1998
Resumen
La proteína quinasa B (PKB) quinasa PDK1 fosforila los isotipos de PKC, específicamente los de PKCzeta y PKCdelta. Esta fosforilación ocurre de una manera dependiente de la fosfoinositida 3-quinasa (PI 3-quinasa), lo que sugiere un papel regulador clave para PDK1 en las vías de señalización de PKC.
Área de la Ciencia:
- Biología Molecular Biología Molecular
- La señalización celular de las células.
- La bioquímica es la bioquímica.
Sus antecedentes:
- Existen sitios de fosforilación conservados en las subfamilias de la proteína quinasa A (PKA), PKG y PKC quinasa.
- La proteína quinasa B (PKB) quinasa, PDK1, es un candidato potencial para la fosforilación de los isotipos de PKC.
Objetivo del estudio:
- Para investigar si la PDK1 fosforila los isotipos de la PKC.
- Para aclarar el mecanismo y la regulación de la fosforilación de PKC mediada por PDK1.
Principales métodos:
- Ensayos de kinasa in vitro utilizando isotipos purificados de PDK1 y PKC.
- Estudios in vivo en células de riñón embrionario humano (293) para evaluar la fosforilación.
- Co-inmunoprecipitación para detectar la formación de complejos proteicos.
- Análisis de la dependencia de la fosfoinositida 3-quinasa (PI 3-quinasa).
Principales resultados:
- PDK1 fosforila directamente los sitios del bucle de activación de PKCzeta y PKCdelta in vitro.
- La fosforilación dependiente de la PDK1 se produjo de una manera dependiente de la PI 3-cinasa in vivo.
- Todos los miembros de la familia PKC probados formaron complejos con PDK1.
- La fosforilación PDK1-dependiente de PKCdelta fue mejorada por los activadores combinados de PKC y PDK1.
- La fosforilación del bucle de activación estimulada por suero de PKCdelta fue dependiente de la PI 3-cinasa y mejorada por la coexpresión de PDK1.
Conclusiones:
- PDK1 es responsable de la fosforilación de PKCzeta y PKCdelta en sus sitios del bucle de activación.
- Esta fosforilación está regulada por la señalización de la PI 3-quinasa.
- PDK1 interactúa con todos los miembros de la familia de PKC probados, lo que indica un papel amplio en la regulación de PKC.
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