Enhancement of odorant detection sensitivity by the expression of odorant-binding protein
Enhancement of odorant detection sensitivity by the expression of odorant-binding protein
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DOI：
10.1016/j.bios.2007.10.008
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发表时间：
2008-02-28
影响因子：
12.6
通讯作者：
Park, Tai Hyun
中科院分区：
文献类型：
--
作者：
Ko, Hwi Jin;Park, Tai Hyun
气味结合蛋白是由鼻腔腺体分泌的低分子量可溶性蛋白。已知它们的功能是运输疏水气味剂。这一特性对于在水相中工作的人工嗅觉生物传感器是重要的。本研究将一种大鼠气味结合蛋白OBP3插入到哺乳动物表达载体pcDNA3中，在人胚胎肾-293 （HEK-293）细胞中表达和分泌。前溶菌酶（plys）的his(6)标签和信号肽分别与OBP3融合检测和分泌蛋白。OBP3分泌量在孵育3 h时达到最大。分泌的OBP3增加了疏水气味剂辛烷醛的溶解度，辛烷醛是大鼠嗅觉受体I7的特异性气味剂。分泌的OBP3也与嗅觉受体I7结合。因此，这些相互作用增加了表达嗅觉受体I7的细胞中气味刺激的细胞信号强度。我们的研究结果表明，气味结合蛋白可以有效地用于提高基于嗅觉受体的生物传感器的灵敏度。(C) 2007 Elsevier B.V.版权所有
Odorant-binding proteins are low molecular weight, soluble proteins that are secreted by glands of the nasal cavity. Their function is known to be the transport of hydrophobic odorants. This feature is important to artificial olfactory biosensors, which operate in the aqueous phase. In this study, one of rat odorant-binding proteins, OBP3, was inserted into a mammalian expression vector pcDNA3, expressed, and secreted from human embryonic kidney-293 (HEK-293) cells. The his(6) tag and signal peptide of the prelysozyme (plys) were fused with OBP3 for the detection and secretion of the proteins, respectively. The secretion level of OBP3 was maximal at 3 h of incubation time. The secreted OBP3 increased the solubility of a hydrophobic odorant, octanal, which is the specific odorant of rat olfactory receptor I7. The secreted OBP3 also bound to olfactory receptor I7. These interactions consequently increased the cellular signal intensity stimulated by the odorant in the cells expressing olfactory receptor I7. Our findings indicate that odorant-binding protein can be effectively used to increase the sensitivity of olfactory receptor-based biosensors. (C) 2007 Elsevier B.V. All rights reserved.