Intracellular trafficking and assembly of specific Kir3 channel/G protein complexes

Localization and Targeting of GIRK Channels in Mammalian Central Neurons

“…RGS7/Gβ5 can be coimmunoprecipitated with GIRK1, GIRK2, and GIRK3 (Xie et al, 2010), as well as with GABA B receptors and other proteins such as R7 family-binding protein (R7BP) (Ostrovskaya et al, 2014). Different Gβγ subunits show a similar ability to activate GIRK channels (Robitaille, Ramakrishnan, Baragli, & Hébert, 2009;Zylbergold et al, 2014). Supporting all these molecular and functional associations, iEM demonstrates that GABA B receptors, GIRK channels, and G and RGS proteins are localized in close proximity at the plasma membrane in different subcellular compartments (Ciruela et al, 2010;Fajardo-Serrano et al, 2013;Kulik et al, 2006).…”

Molecular Pharmacology ofδ-Opioid Receptors

“…Consensus similarly exists with respect to the constitutive association between heterotrimeric G proteins and Kir3 channel effectors (Bünemann et al, 2003;Riven et al, 2006;Robitaille et al, 2009;Berlin et al, 2010Berlin et al, , 2011Richard-Lalonde et al, 2013). Thus, both consensual observations imply that the receptor, G protein, and effector may be part of a single array containing all signaling partners even if for a limited time period, as has been described in HEK cells transfected with DOPrs, Gaob1g2 subunits, and Kir3.1/3.2 channels.…”

Conformational Dynamics of Kir3.1/Kir3.2 Channel Activation Via δ-Opioid Receptors

“…Constructs encoding the yellow fluorescent protein (YFP) or Renilla luciferase (RLuc) fused in frame at the C-terminus of human DORs have been previously described (Breit et al, 2006), as have plasmids encoding YFP fused at the N-terminus of human Gg2 or at the C-terminus of CD8-YFP (Gales et al, 2005). Kir3.1/3.2 human subunits bearing Rluc at their C-terminus have also been described (Robitaille et al, 2009). The recombinant plasmid encoding for GaoA tagged with luciferase at position 99 (GaoA99-Luc) was created from overlapping polymerase chain reaction (PCR) from three fragments, then digested with the restriction enzymes SalI 1 NheI, and cloned in the XhoI 1 NheI sites of the expression vector pCDNA3.1 zeo(1) (Invitrogen, Carlsbad, CA).…”