Cell-based measurement of odorant molecules using surface plasmon resonance

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.

[1]  D. T. Jones,et al.  Biochemical characterization of three stimulatory GTP-binding proteins. The large and small forms of Gs and the olfactory-specific G-protein, Golf. , 1990, The Journal of biological chemistry.

[2]  V. Slepak,et al.  Analysis of protein-protein interactions in phototransduction cascade using surface plasmon resonance. , 2000, Methods in enzymology.

[3]  J. M. McDonnell,et al.  Surface plasmon resonance: towards an understanding of the mechanisms of biological molecular recognition. , 2001, Current opinion in chemical biology.

[4]  Cori Bargmann,et al.  The Caenorhabditis elegans seven-transmembrane protein ODR-10 functions as an odorant receptor in mammalian cells. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[5]  Hiroshi Kataoka,et al.  Molecular Bases of Odor Discrimination: Reconstitution of Olfactory Receptors that Recognize Overlapping Sets of Odorants , 2001, The Journal of Neuroscience.

[6]  Michael L. Shuler,et al.  Growth of endothelial cells on microfabricated silicon nitride membranes for anin vitro model of the blood-brain barrier , 2003 .

[7]  Jeong-Woo Choi,et al.  Nanoscale fabrication of biomolecular layer and its application to biodevices , 2004 .

[8]  T. Z. Wu,et al.  A piezoelectric biosensor as an olfactory receptor for odour detection: electronic nose. , 1999, Biosensors & bioelectronics.

[9]  byung-woo kim,et al.  Application of an interferometric biosensor chip to biomonitoring an endocrine discruptor , 2004 .

[10]  Z. Diénès,et al.  Incorporation of rhodopsin in laterally structured supported membranes: observation of transducin activation with spatially and time-resolved surface plasmon resonance. , 1998, Biochemistry.

[11]  E C Nice,et al.  Instrumental biosensors: new perspectives for the analysis of biomolecular interactions. , 1999, BioEssays : news and reviews in molecular, cellular and developmental biology.

[12]  S. Firestein,et al.  Intracellular trafficking of a tagged and functional mammalian olfactory receptor. , 2002, Journal of neurobiology.

[13]  R. Axel,et al.  A novel multigene family may encode odorant receptors: A molecular basis for odor recognition , 1991, Cell.

[14]  Dietmar Krautwurst,et al.  Identification of Ligands for Olfactory Receptors by Functional Expression of a Receptor Library , 1998, Cell.

[15]  V. Slepak,et al.  Application of surface plasmon resonance for analysis of protein–protein interactions in the G protein‐mediated signal transduction pathway , 2000, Journal of molecular recognition : JMR.

[16]  Hanns Hatt,et al.  Specificity and Sensitivity of a Human Olfactory Receptor Functionally Expressed in Human Embryonic Kidney 293 Cells andXenopus Laevis Oocytes , 1999, The Journal of Neuroscience.

[17]  Cori Bargmann,et al.  A Putative Cyclic Nucleotide–Gated Channel Is Required for Sensory Development and Function in C. elegans , 1996, Neuron.

[18]  Tai Hyun Park,et al.  Piezoelectric olfactory biosensor: ligand specificity and dose-dependence of an olfactory receptor expressed in a heterologous cell system. , 2005, Biosensors & bioelectronics.

[19]  D G Myszka,et al.  Advances in surface plasmon resonance biosensor analysis. , 2000, Current opinion in biotechnology.

[20]  Patrice Mollard,et al.  A single olfactory receptor specifically binds a set of odorant molecules , 2002, The European journal of neuroscience.

[21]  Gregory J Babcock,et al.  Capture and reconstitution of G protein-coupled receptors on a biosensor surface. , 2003, Analytical biochemistry.

[22]  Michihiro Hide,et al.  Real-time analysis of ligand-induced cell surface and intracellular reactions of living mast cells using a surface plasmon resonance-based biosensor. , 2002, Analytical biochemistry.

[23]  Eun Jeong Kim,et al.  Anti-apoptosis engineering , 2003 .

[24]  H. Breer,et al.  Termination of second messenger signaling in olfaction. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[25]  Min-Gon Kim,et al.  Surface plasmon resonance imaging analysis of hexahistidine-tagged protein on the gold thin film coated with a calix crown derivative , 2004 .

[26]  Ikue Mori,et al.  Mutations in a Cyclic Nucleotide–Gated Channel Lead to Abnormal Thermosensation and Chemosensation in C. elegans , 1996, Neuron.

[27]  Cori Bargmann,et al.  odr-10 Encodes a Seven Transmembrane Domain Olfactory Receptor Required for Responses to the Odorant Diacetyl , 1996, Cell.