RESUMO
RGS5 is a R4 type RGS that regulates GPCR signalling. Using western blot, we detected RGS5 as a specific 23 kDa protein in cells overexpressing RGS5. A 42 kDa band representing a possible RGS5 dimer was also detected. Given that GPCRs and their associated proteins form complexes involving multiple protein-protein interactions, we investigated the possibility that the 42 kDa band represents an RGS5-RGS5 dimer. RGS5 dimerization was confirmed by the analysis of a GFP tagged RGS5 fusion in yeast and with two-hybrid assays. Analysis of RGS5 in HEK293A cells suggests that the dimer may serve a regulatory function since it is longer lived than the monomer.
Assuntos
Proteínas RGS/metabolismo , Linhagem Celular , Dimerização , Humanos , Proteínas RGS/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Técnicas do Sistema de Duplo-HíbridoRESUMO
G-protein coupled receptors (GPCRs) represent a large class of cell surface receptors that mediate a multitude of functions. Over the years, a number of GPCRs and ancillary proteins have been shown to be expressed in skeletal muscle. Unlike the case with other muscle tissues like cardiac and vascular smooth muscle cells, there has been little attempt at systematically analyzing GPCRs in skeletal muscle. Here we have compiled all the GPCRs that are expressed in skeletal muscle. In addition, we review the known function of these receptors in both skeletal muscle tissue and in cultured skeletal muscle cells.
Assuntos
Músculo Esquelético/fisiologia , Peptídeos/fisiologia , Receptores Acoplados a Proteínas G/fisiologia , Animais , Proteínas de Transporte/fisiologia , Células Cultivadas , Humanos , Músculo Esquelético/química , Peptídeos/química , Receptores Acoplados a Proteínas G/biossíntese , Transdução de Sinais/fisiologiaRESUMO
Previous investigations had suggested that signaling from the overexpressed beta(2) adrenergic in the heart of transgenic TG4 mice was dampened in the atria. Using an RT-PCR based strategy, we have identified Regulator of G-protein Signaling 5 (RGS5) as being up-regulated in the atria of TG4 mice. Northern blot analysis demonstrated that RGS5 levels were 3 fold higher in the atria of TG4 mice. Western blot analysis of a panel of rat tissues demonstrated that basal expression of RGS5 protein was confined to the heart and skeletal muscle. Furthermore, RGS5 protein was detected in skeletal muscle C2C12 and cardiomyocyte HL-1 cultured cell lines. As observed for RGS5 mRNA levels in TG4 mice, RGS5 protein levels were increased in the atria of rats that were administered the beta adrenergic agonist isoproterenol during a 14 day period. Taken together, these results indicate that RGS5 is a housekeeping RGS in the heart and in skeletal muscle while its beta adrenergic-mediated induction in the atrium suggests that it also has a highly specialized function.
