Piezoelectric biosensor using olfactory receptor protein expressed in Escherichia coli.

An olfactory receptor protein of C. elegans, ODR-10, was expressed in Escherichia coli as a fusion protein, with GST and 6x His-tag. The expression of the target protein was analyzed by SDS-PAGE and Western blot, and was confirmed to be expressed at the membrane fraction of the host E. coli. The surface of a quartz crystal microbalance (QCM) was coated with crude membrane extracts, containing the expressed receptor protein, and the interaction between the olfactory receptor and various odorant molecules examined. Compared with other odorants, diacetyl (2,3-butanedione), known as a natural ligand for the ODR-10 receptor, interacted most strongly with the expressed protein. Various concentrations of diacetyl were applied to the expressed ODR-10 receptor, and the response of the QCM showed a linear relationship to the logarithmic value of the odorant concentration. This piezoelectric biosensor system, using olfactory receptor proteins expressed in E. coli, can be used in diagnostics, toxic chemical detection and the quality control of food.

[1]  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.

[2]  T. Z. Wu,et al.  Synthetic peptide mimicking of binding sites on olfactory receptor protein for use in 'electronic nose'. , 2000, Journal of biotechnology.

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

[4]  R. Vogel,et al.  Bacterial expression of G-protein-coupled receptors: prediction of expression levels from sequence. , 2000, Receptors & channels.

[5]  C. Tate,et al.  Overexpression of integral membrane proteins for structural studies , 1995, Quarterly Reviews of Biophysics.

[6]  L. Buck,et al.  Combinatorial Receptor Codes for Odors , 1999, Cell.

[7]  G. von Heijne,et al.  Properties of N-terminal tails in G-protein coupled receptors: a statistical study. , 1995, Protein engineering.

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

[9]  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.

[10]  H Breer,et al.  Expression of an olfactory receptor in Escherichia coli: purification, reconstitution, and ligand binding. , 1996, Biochemistry.

[11]  D. Lancet Vertebrate olfactory reception. , 1986, Annual review of neuroscience.

[12]  K. Touhara Functional cloning and reconstitution of vertebrate odorant receptors. , 2001, Life sciences.

[13]  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 .

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

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

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

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

[18]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

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

[20]  Gunnar von Heijne,et al.  Fine-tuning the topology of a polytopic membrane protein: Role of positively and negatively charged amino acids , 1990, Cell.