TY - JOUR
T1 - Cell-based measurement of odorant molecules using surface plasmon resonance
AU - Lee, Jin Young
AU - Ko, Hwi Jin
AU - Lee, Sang Hun
AU - Park, Tai Hyun
N1 - Funding Information:
This work was supported by the Korea Science & Engineering Foundation through the Nano-Bioelectronics and Systems Research Center, Seoul National University, Seoul, Korea. We thank Cornelia I. Bargmann (Laboratory of Neural Circuits and Behavior, Howard Hughes Medical Institute, The Rockefeller University, NY, USA) for the odr-10 gene, Randall R. Reed (Department of Neuroscience, Johns Hopkins University, School of Medicine, Baltimore, MD, USA) for the rho-tag gene and Paul A. Hargrave (J. Hillis Miller, Health Center, Department of Ophthalmology, University of Florida College of Medicine, Gainesville, FL, USA) for the B6-30 cell line.
PY - 2006/7/3
Y1 - 2006/7/3
N2 - The surface plasmon resonance (SPR) technique has been used for characterizing molecular interactions. Recently, this technique was used for the analysis of interactions between living cells and molecules reactive to cells. In this study, the technique was applied to the cell-based measurement of odorant molecules. HEK-293 cells were used as a heterologous cell system, and ODR-10, the olfactory receptor of the nematode Caenorhabditis elegans, as a model olfactory receptor. The ODR-10 was expressed on the cell surface, with the help of the rho-tag import sequence, which was fused at the N-terminus of the ODR-10. Exposure of the cells to 0.1 mM diacetyl, which is an odorant molecule specific to the ODR-10, induced a SPR signal from the HEK-293 cells expressing ODR-10, while no SPR signal was detected from the control HEK-293 cells. The intensity of the induced signal was dependent on the dose of diacetyl. These SPR signals were regarded as a result of the intracellular signaling triggered by the binding of odorant molecules to the olfactory receptors. This method, using a heterologous olfactory cell system and SPR system, can be efficiently used to identify the odorant molecules specific to each olfactory receptor.
AB - The surface plasmon resonance (SPR) technique has been used for characterizing molecular interactions. Recently, this technique was used for the analysis of interactions between living cells and molecules reactive to cells. In this study, the technique was applied to the cell-based measurement of odorant molecules. HEK-293 cells were used as a heterologous cell system, and ODR-10, the olfactory receptor of the nematode Caenorhabditis elegans, as a model olfactory receptor. The ODR-10 was expressed on the cell surface, with the help of the rho-tag import sequence, which was fused at the N-terminus of the ODR-10. Exposure of the cells to 0.1 mM diacetyl, which is an odorant molecule specific to the ODR-10, induced a SPR signal from the HEK-293 cells expressing ODR-10, while no SPR signal was detected from the control HEK-293 cells. The intensity of the induced signal was dependent on the dose of diacetyl. These SPR signals were regarded as a result of the intracellular signaling triggered by the binding of odorant molecules to the olfactory receptors. This method, using a heterologous olfactory cell system and SPR system, can be efficiently used to identify the odorant molecules specific to each olfactory receptor.
KW - Diacetyl
KW - HEK-293 cell
KW - Olfactory receptor
KW - Surface plasmon resonance
UR - http://www.scopus.com/inward/record.url?scp=33646821365&partnerID=8YFLogxK
U2 - 10.1016/j.enzmictec.2005.11.036
DO - 10.1016/j.enzmictec.2005.11.036
M3 - Article
AN - SCOPUS:33646821365
SN - 0141-0229
VL - 39
SP - 375
EP - 380
JO - Enzyme and Microbial Technology
JF - Enzyme and Microbial Technology
IS - 3
ER -