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首页> 外文期刊>The Journal of Neuroscience: The Official Journal of the Society for Neuroscience >Specificity and sensitivity of a human olfactory receptor functionally expressed in human embryonic kidney 293 cells and Xenopus Laevis oocytes.
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Specificity and sensitivity of a human olfactory receptor functionally expressed in human embryonic kidney 293 cells and Xenopus Laevis oocytes.

机译：在人类胚胎肾293细胞和非洲爪蟾卵母细胞中功能表达的人类嗅觉受体的特异性和敏感性。

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Here, we provide the first evidence for functional expression of a human olfactory receptor protein (OR17-40) and show that recombinant olfactory receptors can be functionally expressed in heterologous systems. A mixture of 100 different odorants (Henkel 100) elicited a transient increase in intracellular [Ca(2+)] in human embryonic kidney 293 (HEK293) cells stably or transiently transfected with the plasmid pOR17-40. By subdividing the odorant mixture into progressively smaller groups, we identified a single component that represented the only effective substance: helional. Only the structurally closely related molecule heliotroplyacetone also activated the receptor. Other compounds, including piperonal, safrole, and vanillin, were completely ineffective. Mock-transfected cells and cells transfected with other receptors showed no change in intracellular [Ca(2+)] in response to odor stimulation. We were also able to functionally express OR17-40 in Xenopus laevis oocytes. Coexpression of a "reporter" channel allowed measurement of the response of oocytes injected with the cRNA of the human receptor to the odor mixture Henkel 100. The effective substances were the same (helional, heliotropylacetone) as those identified by functionally expressing the receptor in HEK293 cells and were active at the same, lower micromolar concentration. These findings open the possibility of now characterizing the sensitivity and specificity of many, if not all, of the hundreds of different human olfactory receptors.
机译：在这里，我们提供了人类嗅觉受体蛋白（OR17-40）功能表达的第一个证据，并表明重组嗅觉受体可以在异源系统中功能性表达。 100种不同气味的混合物（Henkel 100）在用质粒pOR17-40稳定或瞬时转染的人类胚胎肾293（HEK293）细胞中引起细胞内[Ca（2+）]的瞬时增加。通过将加香剂混合物细分为较小的组，我们确定了代表唯一有效物质的单个成分：螺旋形。只有结构上密切相关的分子巯基丙酮也激活该受体。其他化合物，包括胡椒醛，黄樟脑和香草醛，完全无效。模拟转染的细胞和其他受体转染的细胞对气味刺激的反应没有显示出细胞内[Ca（2+）]的变化。我们还能够在非洲爪蟾卵母细胞中功能性表达OR17-40。 “报告”通道的共表达可以测量注射有人类受体cRNA的卵母细胞对气味混合物汉高100的反应。有效物质与通过在HEK293中功能性表达受体而鉴定出的物质相同（螺旋，巯基丙酮）细胞并在相同的较低微摩尔浓度下具有活性。这些发现打开了现在表征数百种不同的人类嗅觉受体中许多（即使不是全部）敏感性和特异性的可能性。

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