PMID: 7523121Sep 15, 1994Paper

High-level expression of the human neurokinin-1 receptor in mammalian cell lines using the Semliki Forest virus expression system

European Journal of Biochemistry
K LundstromPeter Liljeström

Abstract

Human neurokinin-1 receptor cDNA was introduced into the pSFV1 Semliki Forest virus (SFV) vector and the in vitro transcribed RNA was electroporated into BHK cells with pSFV-Helper RNA. This procedure resulted in the packaging of a high-titer SFV-NK-1 virus stock containing approximately 5 x 10(9) infective units/ml. Infection of baby hamster kidney, COS-7, Chinese hamster ovary and human osteosarcoma cells yielded high levels of human neurokinin-1 receptor expression as assessed by [3H]substance P binding. The maximal receptor expression level obtained was 4 x 10(6) receptors/cell and studies of the post-infection time indicated that a high level of receptor expression was observed 10-24 h post-infection. The human neurokinin-1 receptor expressed in infected baby hamster kidney, COS-7 and Chinese hamster ovary cells was able to stimulate Ca2+ mobilization indicating functional coupling to guanine-nucleotide-binding proteins. The application of the Semliki Forest virus expression system will permit the rapid and efficient production of large quantities of receptor protein for both pharmacological and structural studies.

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Citations

Nov 23, 2006·Journal of Structural and Functional Genomics·Kenneth LundstromFranc Pattus
Oct 1, 1994·Current Opinion in Biotechnology·P Liljeström
Jun 14, 2005·Protein Expression and Purification·Malika AkermounKaty L Gearing
Dec 6, 1997·European Journal of Pharmacology·K LundstromA Naylor
Jun 28, 2000·Journal of the Autonomic Nervous System·W G LachnitA P Ford
Oct 10, 2002·Brain Research Bulletin·Markus U Ehrengruber
Apr 12, 2000·Proceedings of the National Academy of Sciences of the United States of America·F JenckG Kilpatrick
Oct 15, 1996·Proceedings of the National Academy of Sciences of the United States of America·I FrolovC M Rice
Jun 9, 1999·Proceedings of the National Academy of Sciences of the United States of America·M U EhrengruberB H Gähwiler
Oct 1, 1996·The Journal of Cell Biology·B AlletL Bernasconi
Dec 28, 1999·Molecular and Cellular Biology·A SchürmannG M Bokoch
Apr 12, 2013·Future Medicinal Chemistry·Kenneth Lundstrom
Aug 1, 1995·Quarterly Reviews of Biophysics·R Grisshammer, C G Tate
Jan 13, 2009·Journal of Biotechnology·Rim BenmaamarCarlos Augusto Pereira
Jan 22, 2005·Trends in Biotechnology·Kenneth Lundstrom
Mar 15, 2006·Biosensors & Bioelectronics·Simone S KrupkaEva-Kathrin Sinner
Aug 2, 2005·Protein Expression and Purification·Gherici HassaineKenneth Lundstrom
Sep 12, 2012·Molecular Membrane Biology·Juni Andréll, Christopher G Tate
Jan 1, 1995·Journal of Receptor and Signal Transduction Research·K LundstromP Liljestrom
Feb 25, 2003·Molecular Therapy : the Journal of the American Society of Gene Therapy·Kenneth LundstromMarkus U Ehrengruber
Dec 14, 2004·The Journal of Biological Chemistry·Katia MonastyrskayaAnnette Draeger
Apr 11, 2000·Gene Therapy·J J WahlforsR A Morgan
Nov 6, 2001·Journal of Receptor and Signal Transduction Research·K LundstromF Jenck
Dec 31, 2002·Journal of Receptor and Signal Transduction Research·Kenneth Lundstrom
Mar 11, 1999·Journal of Receptor and Signal Transduction Research·K Lundstrom
Oct 6, 2011·Current Protocols in Neuroscience·Markus U EhrengruberKenneth Lundstrom
Jan 1, 1997·Journal of Receptor and Signal Transduction Research·K LundstromA Surprenant

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