Sumoylated RGS-Rz proteins act as scaffolds for Mu-opioid receptors and G-protein complexes in mouse brain.
The RGSZ1 and RGSZ2 proteins, members of the RGS-Rz subfamily of GTPase-activating proteins (GAP), are involved in Mu-opioid receptor desensitization. The expression of these proteins, as well as of their main target the Gz protein, is virtually restricted to the nervous tissue. In synaptosomal membranes, these Rz proteins undergo post-translational modifications such as glycosylation and phosphorylation, and they may covalently attach to small ubiquitin-like modifier (SUMO) proteins. While RGSZ1 exists in conjugated and non-conjugated forms, RGSZ2 is mostly conjugated to SUMO-1, SUMO-2 and SUMO-3 proteins. These sumoylated forms of the GAPs readily associated with Mu-opioid receptors but they associated only poorly with Delta receptors. Furthermore, G alpha i2 and G alpha z subunits co-precipitated with the sumoylated forms of RGSZ1/Z2 proteins, but to a lesser extent with the Ser phosphorylated SUMO-free form of RGSZ1. Upon Mu-opioid receptor activation, there is a strong increase in the association of G alpha proteins with RGSZ2 proteins that persists for intervals longer than 24 h. This effect probably accounts for their role in Mu-opioid receptor desensitization. Only a moderate increase was observed with RGSZ1, the non-sumoylated form of which probably acts as an efficient GAP for these G alpha subunits. Therefore, sumoylation regulates the biological activity of RGS-Rz proteins and it is likely that it serves to switch their behavior, from that of a GAP for activated G alpha subunits to that of a scaffold protein for specific signaling proteins.
Rodríguez-Muñoz
María
M
Department of Neuropharmacology, Cajal Institute, CSIC, Madrid, Spain.
Bermúdez
David
D
Sánchez-Blázquez
Pilar
P
Garzón
Javier
J
D000701:Analgesics, Opioid / Q000494:pharmacology; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D020962:Heterotrimeric GTP-Binding Proteins / Q000378:metabolism*; D047468:Immunoprecipitation / Q000379:methods; D008297:Male; D051379:Mice; D009020:Morphine / Q000494:pharmacology; D010487:Periaqueductal Gray / Q000378:metabolism* / Q000648:ultrastructure; D020710:RGS Proteins / Q000378:metabolism*; D017450:Receptors, Opioid, mu / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D025841:Small Ubiquitin-Related Modifier Proteins / Q000494:pharmacology; D013574:Synaptosomes / Q000187:drug effects / Q000378:metabolism; D013997:Time Factors
D000701:Analgesics, Opioid; D020710:RGS Proteins; C107413:RGS16 protein; D017450:Receptors, Opioid, mu; D025841:Small Ubiquitin-Related Modifier Proteins; D009020:Morphine; D020962:Heterotrimeric GTP-Binding Proteins