Modification of Coding Regions of Target GPCR Genes

One effect of sequence alterations has been to enhance the transcriptional activity that might have a favorable outcome on expression levels. Additionally, deletions of the transmembrane regions have in several cases significantly improved expression levels. In the case of the metabotropic glutamate receptor 1 (mGluR1), the expression of only the extracellular domain generated very high yields and resulted in successful crystallization and a high resolution structure of the domain, which is known to contain the ligand-binding region.1 However, for many GPCRs, the binding pocket is located within one or several of the transmembrane regions. Alternatively, site-directed and random mutagenesis techniques have been applied in attempts to improve the expressability of recombinant proteins.2 Although single point mutations generally resulted in reduced levels or complete loss of expression,3 certain mutations can be advantageous, leading to improved expression levels.4

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