Expression and intracellular localisation of odorant receptors in mammalian cell lines using Semliki Forest virus vectors. 1999

K Monastyrskaia, and F Goepfert, and R Hochstrasser, and G Acuña, and J Leighton, and J R Pink, and K Lundstrom
Givaudan Roure Forschung AG, Dübendorf, Switzerland.

Odorant receptors are members of the G protein-coupled receptor superfamily. They are expressed on the surface of cilia of olfactory neurons, where they bind ligand (odorant). Studies of the molecular mechanisms of olfaction are complicated by the extremely large number of receptor genes, and difficulties in pairing a particular mammalian receptor to a specific odorant ligand in vivo. Here we report expression and localisation studies of two rat odorant receptor genes (17 and OR5), and C. elegans odr-10, using the Semliki Forest virus (SFV) system. All receptors were epitope-tagged at the N- or C-terminus in order to facilitate their detection in infected cells, and determine the localisation and membrane-orientation of recombinant proteins. The immortalised mouse olfactory neuronal cell line OLF 442, rat cortical and striatal primary neuron cultures, and the baby hamster kidney (BHK) cells, were infected and tested. Immunofluorescence and confocal microscopy studies performed on permeabilised, non-permeabilised and native cells revealed that in BHK cells the rat receptors 17 and OR5 were not targeted to the plasma membrane and remained in the endoplasmic reticulum. In contrast, in the mouse olfactory cell line OLF 442 both rat receptors were correctly inserted into the plasma membrane. Similar results were obtained using primary neurons, indicating that like mature neurons, the immortalised OLF 442 cells are capable of providing for correct odorant receptor processing and targeting.

UI MeSH Term Description Entries
D008565 Membrane Proteins Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors. Cell Membrane Protein,Cell Membrane Proteins,Cell Surface Protein,Cell Surface Proteins,Integral Membrane Proteins,Membrane-Associated Protein,Surface Protein,Surface Proteins,Integral Membrane Protein,Membrane Protein,Membrane-Associated Proteins,Membrane Associated Protein,Membrane Associated Proteins,Membrane Protein, Cell,Membrane Protein, Integral,Membrane Proteins, Integral,Protein, Cell Membrane,Protein, Cell Surface,Protein, Integral Membrane,Protein, Membrane,Protein, Membrane-Associated,Protein, Surface,Proteins, Cell Membrane,Proteins, Cell Surface,Proteins, Integral Membrane,Proteins, Membrane,Proteins, Membrane-Associated,Proteins, Surface,Surface Protein, Cell
D009474 Neurons The basic cellular units of nervous tissue. Each neuron consists of a body, an axon, and dendrites. Their purpose is to receive, conduct, and transmit impulses in the NERVOUS SYSTEM. Nerve Cells,Cell, Nerve,Cells, Nerve,Nerve Cell,Neuron
D011994 Recombinant Proteins Proteins prepared by recombinant DNA technology. Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D005822 Genetic Vectors DNA molecules capable of autonomous replication within a host cell and into which other DNA sequences can be inserted and thus amplified. Many are derived from PLASMIDS; BACTERIOPHAGES; or VIRUSES. They are used for transporting foreign genes into recipient cells. Genetic vectors possess a functional replicator site and contain GENETIC MARKERS to facilitate their selective recognition. Cloning Vectors,Shuttle Vectors,Vectors, Genetic,Cloning Vector,Genetic Vector,Shuttle Vector,Vector, Cloning,Vector, Genetic,Vector, Shuttle,Vectors, Cloning,Vectors, Shuttle
D006224 Cricetinae A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS. Cricetus,Hamsters,Hamster
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012672 Semliki forest virus A species of ALPHAVIRUS isolated in central, eastern, and southern Africa.
D013347 Subcellular Fractions Components of a cell produced by various separation techniques which, though they disrupt the delicate anatomy of a cell, preserve the structure and physiology of its functioning constituents for biochemical and ultrastructural analysis. (From Alberts et al., Molecular Biology of the Cell, 2d ed, p163) Fraction, Subcellular,Fractions, Subcellular,Subcellular Fraction
D015801 Helminth Proteins Proteins found in any species of helminth. Helminth Protein,Protein, Helminth,Proteins, Helminth

Related Publications

K Monastyrskaia, and F Goepfert, and R Hochstrasser, and G Acuña, and J Leighton, and J R Pink, and K Lundstrom
January 2010, Methods in molecular biology (Clifton, N.J.),
K Monastyrskaia, and F Goepfert, and R Hochstrasser, and G Acuña, and J Leighton, and J R Pink, and K Lundstrom
August 2001, Biological chemistry,
K Monastyrskaia, and F Goepfert, and R Hochstrasser, and G Acuña, and J Leighton, and J R Pink, and K Lundstrom
May 2001, Current protocols in molecular biology,
K Monastyrskaia, and F Goepfert, and R Hochstrasser, and G Acuña, and J Leighton, and J R Pink, and K Lundstrom
January 1994, Current protocols in molecular biology,
K Monastyrskaia, and F Goepfert, and R Hochstrasser, and G Acuña, and J Leighton, and J R Pink, and K Lundstrom
April 1999, Proceedings of the National Academy of Sciences of the United States of America,
K Monastyrskaia, and F Goepfert, and R Hochstrasser, and G Acuña, and J Leighton, and J R Pink, and K Lundstrom
September 1994, European journal of biochemistry,
K Monastyrskaia, and F Goepfert, and R Hochstrasser, and G Acuña, and J Leighton, and J R Pink, and K Lundstrom
August 2002, Current protocols in human genetics,
K Monastyrskaia, and F Goepfert, and R Hochstrasser, and G Acuña, and J Leighton, and J R Pink, and K Lundstrom
October 2019, Journal of biotechnology,
K Monastyrskaia, and F Goepfert, and R Hochstrasser, and G Acuña, and J Leighton, and J R Pink, and K Lundstrom
August 1999, Insect molecular biology,
K Monastyrskaia, and F Goepfert, and R Hochstrasser, and G Acuña, and J Leighton, and J R Pink, and K Lundstrom
February 2006, Protein expression and purification,
Copied contents to your clipboard!