Heterologous expression of G-protein-coupled receptors: human opioid receptors under scrutiny.
G-protein-coupled receptors whose topology shows seven transmembrane domains form the largest known family of receptors involved in higher organism signal transduction. Despite increasing knowledge on the functioning mechanisms of these receptors, almost no structural data are available but only a few models. Structural studies using a wide range of physical and biochemical techniques may require fairly large (up to several milligrams) amounts of purified protein. Since such quantities are not naturally available, overexpression is prerequisite. Heterologous expression systems are then assayed for maximal production of a protein facsimile. Heterologous systems may also provide interesting alternatives for receptor functional studies in a different cellular context. Opioid receptors will be used as an example to discuss aspects related to the choice and suitability of several different expression systems for the intended analysis of G-protein-coupled receptor properties. General implications will be outlined.
Département des Récepteurs et Protéines membranaires, UPR 9050 CNRS, Ecole supérieure de Biotechnologie de Strasbourg, Illkirch-Graffenstaden, France.
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Stanasila
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Pattus
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Massotte
D000818:Animals; D016367:Baculoviridae; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006801:Humans; D007313:Insecta / Q000166:cytology / Q000821:virology; D008322:Mammals; D009865:Oocytes / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011957:Receptors, Opioid / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D014981:Xenopus; D015003:Yeasts / Q000235:genetics
D011957:Receptors, Opioid; D011994:Recombinant Proteins; D019204:GTP-Binding Proteins