Expression of an olfactory receptor in Escherichia coli: purification, reconstitution, and ligand binding

Biochemistry
Hans KieferH Breer

Abstract

An olfactory receptor has been expressed in bacterial cells as a fusion protein with glutathione S-transferase (GST). Overexpression of receptor protein yielding about 10% of the cell protein was achieved with mutants lacking the N-terminus and the first transmembrane region or with mutants carrying three positively charged residues in the first intracellular loop. The overexpressed fusion protein accumulated in inclusion bodies and could be solubilized in detergent. It was purified by metal chelation chromatography based on a C-terminal 6-histidine tag, and the GST portion was removed after proteolytic cleavage. The purified receptor was reconstituted into lipid vesicles and specific binding of odor ligands was shown by photoaffinity labeling and tryptophan fluorescence measurements. Thus, for the first time, an odorant receptor/ligand pair becomes available in large amounts for biophysical and screening studies.

References

Sep 1, 1992·Bioscience, Biotechnology, and Biochemistry·S MatsuyamaS Mizushima
Dec 1, 1991·Molecular Microbiology·R E Dalbey
Jan 1, 1987·Annals of the New York Academy of Sciences·D LancetU Pace
Apr 5, 1986·Journal of Molecular Biology·R FreudlU Henning
Aug 1, 1995·Quarterly Reviews of Biophysics·R Grisshammer, C G Tate
Jul 1, 1995·Bioconjugate Chemistry·J D OlszewskiG D Prestwich
Apr 1, 1995·Progress in Neurobiology·K Mori, Y Yoshihara
Jan 28, 1993·Nature·K RamingH Breer
Nov 1, 1994·European Journal of Biochemistry·U GatM Natochin
Jan 1, 1994·Annual Review of Biochemistry·C D StraderR A Dixon
Nov 10, 1994·Biochimica Et Biophysica Acta·H BreerJ Krieger
Jan 1, 1994·Critical Reviews in Biochemistry and Molecular Biology·P Pelosi
May 17, 1994·Biochemistry·G Dormán, G D Prestwich
Jul 1, 1995·Protein Engineering·E Wallin, G von Heijne
Aug 1, 1995·Chemical Senses·G D PrestwichS LaForest
May 15, 1996·European Journal of Biochemistry·E NekrasovaU Gat
Aug 1, 1996·The Biochemical Journal·J Tucker, R Grisshammer

❮ Previous
Next ❯

Citations

Mar 1, 2006·Naunyn-Schmiedeberg's Archives of Pharmacology·Daniel BastingClemens Glaubitz
Jul 29, 2009·Analytical and Bioanalytical Chemistry·Claus Hélix Nielsen
Nov 14, 2008·Applied Microbiology and Biotechnology·Jiazhang LianXiaoning Wang
Jun 12, 2009·Applied Microbiology and Biotechnology·Jiazhang LianZhinan Xu
Feb 4, 2009·Environmental Science and Pollution Research International·Ursula Klaschka
Feb 15, 2003·Biochimica Et Biophysica Acta·David DrewJan-Willem L de Gier
Sep 28, 1999·Trends in Pharmacological Sciences·L Dryer, A Berghard
Nov 4, 2000·Biochimica Et Biophysica Acta·M TegoniC Cambillau
Nov 14, 1997·Trends in Biotechnology·I MingarroP Whitley
Jun 27, 1998·Biosensors & Bioelectronics·W GöpelR Malaka
Nov 26, 1998·Biosensors & Bioelectronics·I Lundström, S Svensson
Aug 1, 1997·Current Opinion in Biotechnology·A BohmP B Sigler
Jul 26, 2000·Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology·T Nakamura
May 9, 2012·The Journal of Physical Chemistry. B·Markus EilersSteven O Smith
Sep 21, 2007·Molecular BioSystems·Filippo Mancia, Wayne A Hendrickson
Oct 9, 2008·Proceedings of the National Academy of Sciences of the United States of America·Liselotte KaiserShuguang Zhang
Jul 8, 2009·Proceedings of the National Academy of Sciences of the United States of America·Brian L CookShuguang Zhang
Dec 23, 2009·The Journal of Biological Chemistry·Laure ArcemisbéhèreBernard Mouillac
Sep 18, 2004·European Journal of Biochemistry·Florence VincentChristian Cambillau
May 21, 2013·Journal of Bacteriology·Elaine WoodMaria D Esteve-Gassent
Aug 3, 2010·Biochemistry. Biokhimii︠a︡·L E PetrovskayaM P Kirpichnikov
Apr 2, 2011·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·David N LangelaanJan K Rainey
Sep 27, 2000·Proceedings of the National Academy of Sciences of the United States of America·W B FlorianoG M Shepherd
Mar 31, 1999·Proceedings of the National Academy of Sciences of the United States of America·K TouharaT Haga
May 7, 2011·Annual Review of Biophysics·J-L PopotM Zoonens
Oct 17, 2012·Médecine sciences : M/S·Jean-Louis Banères, Bernard Mouillac
Apr 19, 2011·Trends in Biotechnology·Jean-Louis BanèresBernard Mouillac
Apr 22, 2014·Biochimica Et Biophysica Acta·Bibhuti B DasStanley J Opella
Jul 17, 2010·Progress in Nuclear Magnetic Resonance Spectroscopy·Joseph A GoncalvesSteven O Smith

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.