Journal: The Journal of Neuroscience
Article Title: Glutamate Receptor δ2 Associates with Metabotropic Glutamate Receptor 1 (mGluR1), Protein Kinase Cγ, and Canonical Transient Receptor Potential 3 and Regulates mGluR1-Mediated Synaptic Transmission in Cerebellar Purkinje Neurons
Figure Lengend Snippet: mGluR1, GluRδ2, and PKCγ form protein complexes in cerebellum. A, B, Silver staining of proteins coimmunoprecipitated with mGluR1a or anti-GluRδ2. Each protein lane was excised, divided into six pieces, and analyzed by mass spectroscopy. Proteins identified by their tryptic fragments are indicated. Contaminating IgG, keratin, and trypsin were omitted. C, PKCγ coimmunoprecipitated with mGluR1 or GluRδ2. D, GluRδ2 coimmunoprecipitated with mGluR1 or PKCγ. E, mGluR1a coimmunoprecipitated with GluRδ2 or PKCγ. F, Other metabotropic and ionotropic glutamate receptors did not coimmunoprecipitate with mGluR1, GluRδ2, or PKCγ, confirming the specificity of the IP. G, mGluR1 and GluRδ2 did not associate with other PKC subtypes; H, PKCγ did not associate with mGluR1 or GluRδ2 in cerebral cortex.
Article Snippet: We used following antibodies: rabbit anti-mGluR1a (07-617; Millipore), mouse anti-mGluR1a (clone G209-488; BD Biosciences), rabbit anti-mGluR1a/b (AGC-006; Alamone Labs), anti-GluRδ2 (AB-1514; Millipore; GluRδ2-Rb-Af500-1; Frontier Institute), anti-PKCγ (sc-211; Santa Cruz Biotechnology), anti-PKCα (sc-208; Santa Cruz Biotechnology), anti-PKCβII (sc-210; Santa Cruz Biotechnology), anti-mGluR2/3 (AB1553; Millipore), anti-GluA2 (MAB-397; Millipore), anti-GluN1 (clone 54.1; BD Biosciences Pharmingen), and anti-TRPC3 (ACC-016; Alamone Labs).
Techniques: Silver Staining, Mass Spectrometry