Sushi domains confer distinct trafficking profiles on GABAB receptors.
GABA(B) receptors mediate slow inhibitory neurotransmission in the brain and feature during excitatory synaptic plasticity, as well as various neurological conditions. These receptors are obligate heterodimers composed of GABA(B)R1 and R2 subunits. The two predominant R1 isoforms differ by the presence of two complement control protein modules or Sushi domains (SDs) in the N terminus of R1a. By using live imaging, with an α-bungarotoxin-binding site (BBS) and fluorophore-linked bungarotoxin, we studied how R2 stabilizes R1b subunits at the cell surface. Heterodimerization with R2 reduced the rate of internalization of R1b, compared with R1b homomers. However, R1aR2 heteromers exhibited increased cell surface stability compared with R1bR2 receptors in hippocampal neurons, suggesting that for receptors containing the R1a subunit, the SDs play an additional role in the surface stability of GABA(B) receptors. Both SDs were necessary to increase the stability of R1aR2 because single deletions caused the receptors to be internalized at the same rate and extent as R1bR2 receptors. Consistent with these findings, a chimera formed from the metabotropic glutamate receptor (mGluR)2 and the SDs from R1a increased the surface stability of mGluR2. These results suggest a role for SDs in stabilizing cell surface receptors that could impart different pre- and postsynaptic trafficking itineraries on GABA(B) receptors, thereby contributing to their physiological and pathological roles.
Department of Neuroscience, Physiology, and Pharmacology, University College London, London, United Kingdom.
Saad
S
Hannan
Megan E
ME
Wilkins
Trevor G
TG
Smart
D002038:Bungarotoxins / Q000378:metabolism; D057809:HEK293 Cells; D006624:Hippocampus / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D007091:Image Processing, Computer-Assisted; D018408:Patch-Clamp Techniques; D020033:Protein Isoforms / Q000235:genetics; D055503:Protein Multimerization; D055550:Protein Stability; D017434:Protein Structure, Tertiary / Q000502:physiology; D021122:Protein Subunits / Q000378:metabolism*; D021381:Protein Transport / Q000502:physiology; D018080:Receptors, GABA-B / Q000235:genetics / Q000378:metabolism*; D018094:Receptors, Metabotropic Glutamate / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D009435:Synaptic Transmission / Q000502:physiology*
D002038:Bungarotoxins; D020033:Protein Isoforms; D021122:Protein Subunits; D018080:Receptors, GABA-B; D018094:Receptors, Metabotropic Glutamate; D011993:Recombinant Fusion Proteins; C108822:metabotropic glutamate receptor 2