The basal subcellular distribution of beta-adrenergic receptor kinase is independent of G-protein beta gamma subunits.
beta-Adrenergic receptor kinase (beta ARK-1 or GRK2) is a key regulatory protein involved in the regulation of G-protein-coupled receptors which associates with microsomal and plasma membranes. beta gamma Subunits of G-proteins have been suggested to mediate agonist-dependent membrane translocation of beta ARK, but their possible role in maintaining the complex subcellular distribution of the kinase is not known. In this study we show that lovastatin-mediated inhibition of G gamma subunits isoprenylation in HEK-293 cells stably transfected with beta ARK1 leads to a significant release of G beta subunits to the cytosol without causing changes in total particulate beta ARK or in the association of this kinase to plasma or microsomal membrane fractions. In addition, transient overexpression of mutant forms of G gamma unable to become isoprenylated resulted in a marked sequestration of G beta to the soluble compartment, but caused no rearrangement in the distribution of cotransfected beta ARK. These results indicate that anchoring of beta ARK to cellular membranes under basal conditions is independent of the availability of heterotrimeric G-protein subunits.
Murga
C
C
Departamento de Biología Molecular, Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Universidad Autónoma de Madrid, Spain.
Esteban
N
N
Ruiz-Gómez
A
A
Mayor
F
F
D002460:Cell Line; D017868:Cyclic AMP-Dependent Protein Kinases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D006801:Humans; D007425:Intracellular Membranes / Q000187:drug effects / Q000201:enzymology; D007668:Kidney; D008148:Lovastatin / Q000494:pharmacology; D017368:Protein Prenylation; D013347:Subcellular Fractions / Q000187:drug effects / Q000201:enzymology; D051552:beta-Adrenergic Receptor Kinases
D008148:Lovastatin; D017868:Cyclic AMP-Dependent Protein Kinases; D051552:beta-Adrenergic Receptor Kinases; D019204:GTP-Binding Proteins