Tamalin, a PDZ Domain-Containing Protein, Links a Protein Complex Formation of Group 1 Metabotropic Glutamate Receptors and the Guanine Nucleotide Exchange Factor Cytohesins

• Published in: The Journal of neuroscience Vol. 22; no. 4; pp. 1280 - 1289
• Main Authors: Kitano, Jun, Kimura, Kouji, Yamazaki, Yoshimitsu, Soda, Takeshi, Shigemoto, Ryuichi, Nakajima, Yoshiaki, Nakanishi, Shigetada
• Format: Journal Article
• Language: English
• Published: United States Soc Neuroscience 15.02.2002; Society for Neuroscience
• Subjects: ADP-Ribosylation Factors - metabolism; Animals; Brain Chemistry; Carrier Proteins - genetics; Carrier Proteins - metabolism; Cell Adhesion Molecules - metabolism; Cells, Cultured; Cloning, Molecular; COS Cells; Guanine Nucleotide Exchange Factors - metabolism; Humans; In Situ Hybridization; Macromolecular Substances; Membrane Proteins - genetics; Membrane Proteins - metabolism; Mice; Molecular Sequence Data; Neurons - cytology; Neurons - metabolism; Organ Specificity; Precipitin Tests; Protein Structure, Tertiary - physiology; Protein Transport - drug effects; Protein Transport - physiology; Rats; Receptor, Metabotropic Glutamate 5; Receptors, Metabotropic Glutamate - metabolism; RNA, Messenger - metabolism; Sequence Homology, Amino Acid; Transfection; Two-Hybrid System Techniques
• ISSN: 0270-6474; 1529-2401
• DOI: 10.1523/jneurosci.22-04-01280.2002

In this investigation, we report identification and characterization of a 95 kDa postsynaptic density protein (PSD-95)/discs-large/ZO-1 (PDZ) domain-containing protein termed tamalin, also recently named GRP1-associated scaffold protein (GRASP), that interacts with group 1 metabotropic glutamate receptors (mGluRs). The yeast two-hybrid system and in vitro pull-down assays indicated that the PDZ domain-containing, amino-terminal half of tamalin directly binds to the class I PDZ-binding motif of group 1 mGluRs. The C-terminal half of tamalin also bound to cytohesins, the members of guanine nucleotide exchange factors (GEFs) specific for the ADP-ribosylation factor (ARF) family of small GTP-binding proteins. Tamalin mRNA is expressed predominantly in the telencephalic region and highly overlaps with the expression of group 1 mGluR mRNAs. Both tamalin and cytohesin-2 were enriched and codistributed with mGluR1a in postsynaptic membrane fractions. Importantly, recombinant and native mGluR1a/tamalin/cytohesin-2 complexes were coimmunoprecipitated from transfected COS-7 cells and rat brain tissue, respectively. Transfection of tamalin and mutant tamalin lacking a cytohesin-binding domain caused an increase and decrease in cell-surface expression of mGluR1a in COS-7 cells, respectively. Furthermore, adenovirus-mediated expression of tamalin and dominant-negative tamalin facilitated and reduced the neuritic distribution of endogenous mGluR5 in cultured hippocampal neurons, respectively. The results indicate that tamalin plays a key role in the association of group 1 mGluRs with the ARF-specific GEF proteins and contributes to intracellular trafficking and the macromolecular organization of group 1 mGluRs at synapses.