Solubilization of A1 adenosine receptor from pig brain: characterization and evidence of the role of the cell membrane on the coexistence of high- and low-affinity states

Journal of Neuroscience Research
Vicent CasadóR Franco

Abstract

The present solubilization strategy recognizes the important role of detergent cocktails in the solubilization and subsequent stability of adenosine A1, receptors from pig brain cortical membranes. The 3-[3-(cholamidopropyl)dimethylammonio]-1-propane-sulfonate-digitonin mixture produced the extraction of up to 52% of the receptor with an enrichment of 1.2-fold with respect to crude membranes. The binding activity of the soluble extract was very stable even in the absence of glycerol. In crude membranes the existence of high- and low-affinity states was detected, but in the soluble extract and in the detergent-treated membranes only the high-affinity state was detected. Association-dissociation curves showed that in crude membranes no interconversion between high- and low-affinity sites is produced by the association of the ligand [3H]R-N6-phenylisopropyladenosine. These results suggest that the high- and low-affinity states are different conformations induced by the structure of the membrane. The modulation of the binding activity by (Gpp(NH)p) 5'-guanylylimidodiphosphate and Mg2+ was studied. In crude membranes Gpp(NH)p shifted the high-affinity state to the low-affinity state, whereas the contrary occurred when Mg2+ was used....Continue Reading

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Citations

Jul 1, 1997·Progress in Neurobiology·R FrancoC Lluis
Mar 30, 2002·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Joseph F Siebenaller, Deiadra J Garrett
May 20, 2006·Neuropsychopharmacology : Official Publication of the American College of Neuropsychopharmacology·Davide QuartaSergi Ferré
Nov 1, 1992·British Journal of Pharmacology·J BlancoR Franco
Apr 29, 1998·Immunological Reviews·R FrancoJ Blanco
Jan 25, 2005·Acta Pharmacologica Sinica·Wan-Chun SunXing-Zu Zhu
Apr 1, 2010·Expert Opinion on Drug Discovery·Rafael FrancoSergi Ferre
Oct 11, 2015·Molecular Neurobiology·Rafael FrancoVicent Casadó
Nov 27, 1991·Biochemical and Biophysical Research Communications·G AllendeE I Canela
Oct 26, 2014·Molecular Neurobiology·Daniel FarréRafael Franco
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Dec 15, 1995·Journal of Neuroscience Research·F CiruelaR Franco
May 6, 2009·British Journal of Pharmacology·Carla FerradaRafael Franco
May 19, 2010·Journal of Neurochemistry·Vicent CasadóRafael Franco
Feb 28, 2006·Molecular Pharmacology·Rafael FrancoEnric I Canela
Dec 23, 2009·The Journal of Pharmacology and Experimental Therapeutics·Aroa SorianoVicent Casadó
Nov 30, 2012·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Eduard GraciaVicent Casadó
Mar 14, 2019·Biomolecular Concepts·Rafael FrancoGemma Navarro
Jun 2, 2007·Journal of Medicinal Chemistry·Marc VendrellMiriam Royo

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