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Functional Analysis of Olfactory Receptors Expressed in a HEK-293 Cell System by Using Cameleons  

Ko, Hwi-Jin (School of Chemical and Biological Engineering, Seoul National University)
Park, Tai-Hyun (School of Chemical and Biological Engineering, Seoul National University)
Publication Information
Journal of Microbiology and Biotechnology / v.17, no.6, 2007 , pp. 928-933 More about this Journal
Abstract
Cameleon is a genetically engineered $Ca^{2+}$ sensing molecule consisting of two variants of the green fluorescent protein (GFP), calmodulin and calmodulin-binding protein, M13. HEK-293 cells stably expressing three types of cameleons, yellow cameleon-2, cameleon-3er, and cameleon-2nu, were constructed, and the expression and localization of these cameleons were confirmed by fluorescent imaging. Among the cameleons, the yellow cameleon-2 was selected for analyzing the change in $Ca^{2+}$ induced by the olfactory receptor-mediated signal transduction, because it is localized in the cytosol and binds to cytosolic $Ca^{2+}$ ions. Cells stably expressing yellow cameleon-2 were transfected with each of the test olfactory receptor genes, odr-10 and 17, and the expression of the olfactory receptor genes were examined using immunocytochenmical methods and RT-PCR. Stimulating each olfactory receptor with its specific odorant caused an increase in the intracellular $Ca^{2+}$ level, which was measured using yellow cameleon-2. These results demonstrate that yellow cameleon-2 can be conveniently used to examine the function of the olfactory receptors expressed in heterologous cells.
Keywords
Cameleon; olfactory receptor; HEK-293 cells; $Ca^{2+}$;
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Times Cited By KSCI : 5  (Citation Analysis)
Times Cited By Web Of Science : 4  (Related Records In Web of Science)
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