Abstract
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 his6 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.
Original language | English |
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Pages (from-to) | 1017-1023 |
Number of pages | 7 |
Journal | Biosensors and Bioelectronics |
Volume | 23 |
Issue number | 7 |
DOIs | |
State | Published - 28 Feb 2008 |
Bibliographical note
Funding Information:This work was supported by the Korea Science & Engineering Foundation through the Nano-Bioelectronics and Systems Research Center, Seoul National University, Seoul, Republic of Korea. We thank Heinz Breer (Institute of Physiology, University Hohenheim, Stuttgart, Germany) for the OBP3 gene, Randall R. Reed (Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA) for the rho-tag gene, and Akihito Yasuoka (Bio-oriented Technology Research Advancement Institution, Saitama, Japan) for the pVL-I7 .
Keywords
- HEK-293
- Odorant-binding protein
- Olfactory receptor I7
- Olfactory receptor-based olfactory biosensor