A new technique to co-localise membrane proteins with Homer/vesl

作者：Hiroaki Y; Nishikawa K; Mitsuoka K; Tachibana T; Sobue K; Doi T; Fujiyoshi Y^*

来源：Biochemical and Biophysical Research Communications, 2002, 295(3): 756-765.

DOI：10.1016/S0006-291X(02)00738-6

• endothelin B receptor
• metabotropic glutamate receptor
• G-protein-coupled receptors
• Homer 1c
• proline-rich
• localisation
• 2D-crystallisation
• co-expression
• electron crystallography
• postsynapse

The minimal requirements were defined as necessary for cluster formation of the group I 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 (ETBRs). 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 ETBR/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.

• 出版日期2002-7-19