Functional G-Protein-Coupled Receptor (GPCR) Synthesis: The Pharmacological Analysis of Human Histamine H1 Receptor (HRH1) Synthesized by a Wheat Germ Cell-Free Protein Synthesis System Combined with Asolectin Glycerosomes

Frontiers in Pharmacology
Yasuyuki SuzukiKazutaka Maeyama

Abstract

G-protein-coupled receptors (GPCRs) are membrane proteins distributed on the cell surface, and they may be potential drug targets. However, synthesizing GPCRsin vitrocan be challenging. Recently, some cell-free protein synthesis systems have been shown to produce a large amount of membrane protein combined with chemical chaperones that include liposomes and glycerol. Liposomes containing high concentrations of glycerol are known as glycerosomes, which are used in new drug delivery systems. Glycerosomes have greater morphological stability than liposomes. Proteoglycerosomes are defined as glycerosomes that contain membrane proteins. Human histamine H1receptor (HRH1) is one of the most studied GPCRs. In this study, we synthesized wild-type HRH1 (WT-HRH1) proteoglycerosomes and D107A-HRH1, (in which Asp107 was replaced by Ala) in a wheat germ cell-free protein synthesis system combined with asolectin glycerosomes. The mutant HRH1 has been reported to have low affinity for the H1antagonist. In this study, the amount of synthesized WT-HRH1 in one synthesis reaction was 434 ± 66.6 μg (7.75 ± 1.19 × 103pmol). The specific binding of [3H]pyrilamine to the WT-HRH1 proteoglycerosomes became saturated as the concentration of the radioliga...Continue Reading

References

Sep 15, 1994·Biochemical and Biophysical Research Communications·K OhtaH Fukui
Oct 9, 1999·The Journal of Biological Chemistry·K WielandR Leurs
Oct 16, 1999·Science·R L StrausbergF S Collins
Jan 19, 2000·Proceedings of the National Academy of Sciences of the United States of America·K MadinY Endo
Mar 23, 2004·European Journal of Pharmacology·Martijn BruystersRob Leurs
Jun 23, 2004·European Journal of Biochemistry·Venkata R P RatnalaWillem J DeGrip
Jun 29, 2004·Journal of Thrombosis and Haemostasis : JTH·S A Smith, J H Morrissey
Mar 14, 2007·Journal of Structural Biology·Christian KlammtFrank Bernhard
Jul 20, 2007·Nature Chemical Biology·James IngleseDouglas S Auld
Sep 29, 2007·Journal of Biotechnology·Shin-Ichiro M NomuraKazunari Akiyoshi
Apr 3, 2008·Nature Reviews. Drug Discovery·Malin C Lagerström, Helgi B Schiöth
Jul 21, 2009·Trends in Biotechnology·Federico KatzenWieslaw Kudlicki
Sep 12, 2009·Protein Science : a Publication of the Protein Society·Kazumi ShimonoShigeyuki Yokoyama
Feb 4, 2010·PloS One·Isabelle Mouro-ChanteloupPierre Ripoche
Feb 6, 2010·Nature Protocols·Kazuyuki TakaiYaeta Endo
Apr 6, 2011·Protein Science : a Publication of the Protein Society·Christian KlammtSenyon Choe
Jun 24, 2011·Nature·Tatsuro ShimamuraSo Iwata
Feb 7, 2012·Journal of Molecular Biology·Sayaka InagakiReinhard Grisshammer
Jun 22, 2012·Molecular Membrane Biology·Christian RoosFrank Bernhard
Aug 6, 2013·International Journal of Pharmaceutics·Maria Letizia MancaAnna Maria Fadda
Jun 17, 2014·FEBS Letters·Matthias Harbers
Jun 19, 2015·Pharmacological Reviews·Pertti PanulaHelmut L Haas
Nov 17, 2015·Nature Chemical Biology·Rosie DawalibyCédric Govaerts
Jan 23, 2016·International Journal of Molecular Sciences·Yinfeng YangLing Yang
Jul 21, 2016·European Biophysics Journal : EBJ·Ida Louise JørgensenThomas Günther Pomorski
Feb 9, 2017·Chemical Reviews·Ilia G Denisov, Stephen G Sligar
Jun 3, 2017·Biotechnology and Bioengineering·Andrei SonnabendStefan Kubick
Jun 18, 2017·Scientific Reports·Anne ZemellaStefan Kubick

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Citations

Sep 21, 2019·Biological Chemistry·Lisa Maria KöglerAnnette G Beck-Sickinger
Mar 22, 2020·BioDrugs : Clinical Immunotherapeutics, Biopharmaceuticals and Gene Therapy·Srujan Kumar DondapatiStefan Kubick
Oct 21, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Andrew J Y JonesDaniel Nietlispach

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Methods Mentioned

BETA
protein folding
PCR
electrophoresis
density gradient centrifugation
X-ray
Protein Assay
saturation binding

Software Mentioned

ImageJ
EZR
GraphPad
Prism
GraphPad Prism

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