A new technique to co-localise membrane proteins with Homer/vesl |
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Authors: | Hiroaki Yoko Nishikawa Kouki Mitsuoka Kaoru Tachibana Taro Sobue Kenji Doi Tomoko Fujiyoshi Yoshinori |
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Affiliation: | Department of Biophysics, Graduate School of Science, Kyoto University, Oiwake, Kitashirakawa, Sakyo-ku, Japan. |
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Abstract: | The minimal requirements were defined as necessary for cluster formation of the group 1 metabotropic glutamate receptor (mGluR), which is regulated by the Homer/vesl family of scaffolding proteins [Curr. Opin. Neurobiol. 10 (2000) 370]. Cluster formation of G-protein-coupled receptors (GPCRs) plays a fundamental role in signal transduction, particularly at the neuronal synapse. To understand the interaction of mGluR with PSD-Zip45, a Homer/vesl family member, we designed a series of chimeric receptor proteins, consisting of C-terminal mGluR1alpha sequences that were fused to endothelin B receptors (ET(B)Rs). In vitro and in vivo studies revealed that an extended 20 amino acid long C-terminal mGluR1alpha peptide, including the proline-rich core motif PPXXF, is sufficient to induce clustering of chimeric ET(B)R/mGluR1alpha receptors by PSD-Zip45. This result is especially important because it constitutes the basis for a new approach to form two-dimensional crystals of membrane proteins in situ, which may render unstable membrane proteins amenable to electron crystallographic structure determination. |
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Keywords: | Endothelin B receptor Metabotropic glutamate receptor G-protein-coupled receptors Homer 1c Proline-rich Localisation 2D-crystallisation Co-expression Electron crystallography Postsynapse |
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