Mammalian expression of transmembrane receptors for pharmaceutical applications.
Three mammalian expression systems suitable for expressing recombinant receptors have been described. Each is suited to a different aspect of the study of receptors and their behaviour. IRES-based vectors are ideal for creating stable mammalian cell lines suitable for screening receptors using a signalling readout. Unlike traditional vectors they result in almost 100% of cell lines generated expressing a particular receptor, thus increasing the efficiency of cell line generation and increasing the chance of higher expression-level cell lines being generated. They may also be utilized to express more than one protein of interest, for example it is possible to co-express a particular receptor with a particular signalling protein or trafficking protein from a single RNA, thus ensuring that both are expressed simultaneously in the same cell. The ecdysone-inducible expression system is ideal for studying receptor signalling and behaviour. It is possible to alter receptor expression levels in an identical cellular background thus making it possible to study phenomena such as constitutive receptor activity in the absence of agonist. The SFV expression system is ideal for expressing receptors at high levels of a mammalian cell. It is thus a good system for purifying receptors for structural analysis and for providing material for binding assays. All of the expression systems described above have been demonstrated to express seven-transmembrane receptors with the expected pharmacological and functional profile.
Receptor Systems Unit, Glaxo Wellcome Medicines Research Centre, Stevenage, UK.
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Rhodes
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Bevan
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Patel
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Lee
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Rees
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D015195:Drug Design; D005822:Genetic Vectors; D006801:Humans; D011956:Receptors, Cell Surface / Q000235:genetics*
D011956:Receptors, Cell Surface