title: Development and characterization of the α3β4α5 nicotinic receptor cellular membrane affinity chromatography column and its application for on line screening of plant extracts.
abstract: The α3β4α5 nAChR has been recently shown to be a useful target for smoking cessation pharmacotherapies. Herein, we report on the development and characterization of the α3β4α5 nicotinic receptor column by frontal displacement chromatography. The binding affinity of the nicotine and minor alkaloids found in tobacco smoke condensates were determined for both the α3β4 and α3β4α5 nicotinic receptors. It was demonstrated that while no subtype selectivity was observed for nicotine and nornicotine, anabasine was selective for the α3β4α5 nicotinic receptor. The non-competitive inhibitor binding site was also studied and it was demonstrated while mecamylamine was not selective between subtypes, buproprion showed subtype selectivity for the α3β4 nicotinic receptor. The application of this methodology to complex mixtures was then carried out by screening aqueous-alcoholic solutions of targeted plant extracts, including Lycopodium clavatum L. (Lycopodiaceae) and Trigonella foenum graecum L. (Fabaceae) against both the α3β4 and α3β4α5 nAChRs.
authors.affiliation: Laboratory of Clinical Investigation, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, United States; Department of Inorganic Chemistry, Medical University of Lublin, Chodzki 4A, 20-093 Lublin, Poland.
authors.forename: L
authors.initials: L
authors.lastname: Ciesla
authors.affiliation: Laboratory of Clinical Investigation, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, United States.
authors.forename: M
authors.initials: M
authors.lastname: Okine
authors.affiliation: Laboratory of Clinical Investigation, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, United States.
authors.forename: A
authors.initials: A
authors.lastname: Rosenberg
authors.affiliation: Laboratory of Clinical Investigation, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, United States.
authors.forename: K S S
authors.initials: KSS
authors.lastname: Dossou
authors.affiliation: Torrey Pines Institute for Molecular Studies, Port St. Lucie, FL 34990, United States.
authors.forename: L
authors.initials: L
authors.lastname: Toll
authors.affiliation: Laboratory of Clinical Investigation, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, United States.
authors.forename: I W
authors.initials: IW
authors.lastname: Wainer
authors.affiliation: Laboratory of Clinical Investigation, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, United States. Electronic address: moaddelru@mail.nih.gov.
authors.forename: R
authors.initials: R
authors.lastname: Moaddel
mesh_terms: D000470:Alkaloids / Q000737:chemistry; D000691:Anabasine / Q000737:chemistry; D001665:Binding Sites; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002846:Chromatography, Affinity; D007887:Fabaceae / Q000737:chemistry; D029542:Lycopodiaceae / Q000737:chemistry; D009538:Nicotine / Q000031:analogs & derivatives / Q000737:chemistry; D010936:Plant Extracts / Q000737:chemistry*; D011978:Receptors, Nicotinic / Q000737:chemistry*; D012906:Smoke / Q000032:analysis
chemical_list: D000470:Alkaloids; D010936:Plant Extracts; D011978:Receptors, Nicotinic; D012906:Smoke; C403530:nicotinic receptor alpha3beta4; D009538:Nicotine; C005311:nornicotine; D000691:Anabasine
keywords: Alkaloids; Anabasine; Frontal affinity chromatography; Subtype selectivity