Mapping the isoprenoid binding pocket of PDEdelta by a semisynthetic, photoactivatable N-Ras lipoprotein.
Chembiochem
; 10(1): 98-108, 2009 Jan 05.
Article
en En
| MEDLINE
| ID: mdl-18846587
ABSTRACT
Biologically functional Ras isoforms undergo post-translational modifications starting with farnesylation of the most C-terminal cysteine. Combined with further processing steps, this isoprenylation allows for the anchoring of these proteins in endomembranes, where signal transduction events take place. The specific localization is subject to dynamic regulation and assumed to modulate the activity of Ras proteins by governing their spatiotemporal distribution. The delta subunit of phosphodiesterase (PDEdelta) has attracted attention as a solubilization factor of isoprenylated Ras. In this study, we demonstrate that critical residues in the putative isoprenoid pocket of PDEdelta can be mapped by coupling with a semisynthetic N-Ras lipoprotein in which the native farnesyl group of the processed protein was replaced by a photoactivatable geranyl benzophenone moiety. The crosslinked product included parts of beta-sheet 9 of PDEdelta, which contains the highly conserved amino acids V145 and L147. Modeling of the PDEdelta-geranyl benzophenone (GerBP) complex supports the conclusion that the photolabeled sequence is embedded in the putative isoprenoid pocket of PDEdelta.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Terpenos
/
Hidrolasas Diéster Fosfóricas
/
Proteínas ras
/
Luz
/
Lipoproteínas
Tipo de estudio:
Prognostic_studies
Límite:
Humans
Idioma:
En
Año:
2009
Tipo del documento:
Article