title: Uncovering the lipidic basis for the preparation of functional nicotinic acetylcholine receptor detergent complexes for structural studies.
abstract: This study compares the lipid composition, including individual phospholipid molecular species of solubilized nAChR detergent complexes (nAChR-DCs) with those of the bulk lipids from their source, Torpedo californica (Tc) electric tissue. This lipidomic analysis revealed seventy-seven (77) phospholipid species in the Tc tissue. Analysis of affinity-purified nAChR-DCs prepared with C-12 to C-16 phospholipid analog detergents alkylphosphocholine (FC) and lysofoscholine (LFC) demonstrated that nAChR-DCs prepared with FC12, LFC14, and LFC16 contained >60 phospholipids/nAChR, which was more than twice of those prepared with FC14, FC16, and LFC12. Significantly, all the nAChR-DCs lacked ethanolamine and anionic phospholipids, contained only four cholesterol molecules, and a limited number of phospholipid molecular species per nAChR. Upon incorporation into oocytes, FC12 produce significant functionality, whereas LFC14 and LFC16 nAChR-DCs displayed an increased functionality as compared to the crude Tc membrane. All three nAChR-DCs displayed different degrees of alterations in macroscopic activation and desensitization kinetics.
authors.affiliation: Department of Physical Sciences, University of Puerto Rico, Río Piedras Campus, PO Box 23323, San Juan, 00931-03323, Puerto Rico.
authors.forename: Orestes
authors.initials: O
authors.lastname: Quesada
authors.affiliation: Department of Chemistry, University of Puerto Rico, Río Piedras Campus, PO Box 23346, San Juan, 00931-3346, Puerto Rico.
authors.forename: Carol
authors.initials: C
authors.lastname: González-Freire
authors.affiliation: Department of Biology, University of Puerto Rico, Río Piedras Campus, PO Box 70377, San Juan, 00936-8377, Puerto Rico.
authors.forename: María Carla
authors.initials: MC
authors.lastname: Ferrer
authors.affiliation: Department of Pharmaceutical Sciences, School of Pharmacy, University of Puerto Rico, San Juan, 00936, Puerto Rico.
authors.forename: José O
authors.initials: JO
authors.lastname: Colón-Sáez
authors.affiliation: Department of Biology, University of Puerto Rico, Río Piedras Campus, PO Box 70377, San Juan, 00936-8377, Puerto Rico.
authors.forename: Emily
authors.initials: E
authors.lastname: Fernández-García
authors.affiliation: Department of Biology, University of Puerto Rico, Río Piedras Campus, PO Box 70377, San Juan, 00936-8377, Puerto Rico.
authors.forename: Juan
authors.initials: J
authors.lastname: Mercado
authors.affiliation: Department of Biology, University of Puerto Rico, Río Piedras Campus, PO Box 70377, San Juan, 00936-8377, Puerto Rico.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Dávila
authors.affiliation: Department of Chemistry, University of Puerto Rico, Río Piedras Campus, PO Box 23346, San Juan, 00931-3346, Puerto Rico.
authors.forename: Reginald
authors.initials: R
authors.lastname: Morales
authors.affiliation: Department of Chemistry, University of Puerto Rico, Río Piedras Campus, PO Box 23346, San Juan, 00931-3346, Puerto Rico.|Department of Biology, University of Puerto Rico, Río Piedras Campus, PO Box 70377, San Juan, 00936-8377, Puerto Rico.|University of Puerto Rico, Molecular Sciences and Research Center, San Juan, 00926, Puerto Rico.
authors.forename: José A
authors.initials: JA
authors.lastname: Lasalde-Dominicci
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D002384:Catalysis; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D003902:Detergents / Q000737:chemistry*; D004557:Electric Organ / Q000378:metabolism; D004566:Electrodes; D006868:Hydrolysis; D008055:Lipids / Q000737:chemistry* / Q000302:isolation & purification; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism; D018408:Patch-Clamp Techniques; D011978:Receptors, Nicotinic / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D014101:Torpedo; D014981:Xenopus
chemical_list: D003902:Detergents; D008055:Lipids; D011978:Receptors, Nicotinic; D000109:Acetylcholine