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Biophysical and Ion Channel Functional Characterization of the Torpedo Californica Nicotinic Acetylcholine Receptor in Varying Detergent-Lipid Environments
dc.contributor.author | Asmar-Rovira, Guillermo A. | |
dc.contributor.author | Asseo-García, Aloysha M. | |
dc.contributor.author | Quesada, Orestes | |
dc.contributor.author | Hanson, Michael A. | |
dc.contributor.author | Nogueras, Carlos | |
dc.contributor.author | Lasalde-Dominicci, José A. | |
dc.contributor.author | C. Stevens, Raymond | |
dc.date.accessioned | 2017-05-31T21:02:42Z | |
dc.date.available | 2017-05-31T21:02:42Z | |
dc.date.copyright | All of the material available from the PMC site is provided by the respective publishers or authors. Almost all of it is protected by U.S. and/or foreign copyright laws, even though PMC provides free access to it. (See Public Domain Material below, for one exception.) The respective copyright holders retain rights for reproduction, redistribution and reuse. Users of PMC are directly and solely responsible for compliance with copyright restrictions and are expected to adhere to the terms and conditions defined by the copyright holder. Transmission, reproduction, or reuse of protected material, beyond that allowed by the fair use principles of the copyright laws, requires the written permission of the copyright owners. U.S. fair use guidelines are available from the Copyright Office at the Library of Congress. | en_US |
dc.date.issued | 2009-05-01 | |
dc.identifier.citation | Asmar-Rovira, G. A., Asseo-García, A. M., Quesada, O., Hanson, M. A., Nogueras, C., Lasalde-Dominicci, J. A., & Stevens, R. C. (2008). Biophysical and ion channel functional characterization of the Torpedo californica nicotinic acetylcholine receptor in varying detergent-lipid environments. The Journal of Membrane Biology, 223(1), 13–26. http://doi.org/10.1007/s00232-008-9107-7 | en_US |
dc.identifier.issn | 1432-1424 | |
dc.identifier.uri | http://hdl.handle.net/11721/1623 | |
dc.description.abstract | The nicotinic acetylcholine receptor (nAChR) of Torpedo electric rays has been extensively characterized over the last three decades. However, the molecular mechanisms by which detergents influence membrane protein stability and function remain poorly understood, and elucidation of the dynamic detergent-lipid-protein interactions of solubilized membrane proteins is a largely unexplored research field. This study examined nine detergents upon nAChR solubilization and purification, to assess receptor lipid composition using GC (Gas Chromatography)-FID (Flame Ionization) and/or GC-MSD (Mass Selective Detectors), stability and aggregation state using A-SEC (Analytical Size-Exclusion Chromatography) and EM (Electron Microscopy), and planar lipid bilayers to measure ion channel function. Detergent solubilization of nAChR-enriched membranes did not result in significant native lipid depletion or destabilization. Upon purification, native lipid depletion occurred in all detergents, with lipid-analog detergents [CHAPS (3-[(3-Cholamidopropyl)-dimethylammonio]-1-propane sulfonate), FC-12 (n-Dodecylphosphocholine) and sodium cholate (3α,7α,12α-Trihydroxy-5β-cholan-24-oic acid)] maintaining stability and supporting ion channel function, while non-lipid analog detergents [Cymal-6 (6-Cyclohexyl-1-hexyl-β-d-maltoside), DDM (n-Dodecyl-β-d-maltopyranoside), LDAO (Lauryldimethylamine-N-oxide) and OG (n-Octyl-β-d-glucopyranoside)] showed decreased stability and significant reduction or loss of ion channel function. Anapoe-C12E9 (Polyoxyethylene-(9)-dodecyl ether) and BigCHAP (N,N′-bis-(3-d-Gluconamidopropyl) cholamide) retained residual amounts of native lipid, maintaining moderate stability and ion channel function when compared to lipid-analog detergents. Overall, these results show that the nAChR can be stable and functional in lipid-analog detergents or in detergents that retain moderate amounts of residual native lipids, while the opposite is true about non-lipid analog detergents. These results highlight the importance of careful biophysical characterization of membrane proteins for future functional or structural studies in the detergent-solubilized state. | en_US |
dc.description.sponsorship | NIH (National Institutes of Health) grants R01GM056371, S06GM008102, SNRP U54NS043011 and UPR Institutional Funds for Research (to José A. Lasalde-Dominicci), the NIH-MBRS (Minority Biomedical Research Support) Research Initiative for Scientific Enhancement (RISE) grant R25GM061151 and the NIH Roadmap grant P50GM073197 (to Raymond C. Stevens) and Alliances for Graduate Education in the Professoriate (AGEP) and RISE fellowships (to Guillermo A. Asmar-Rovira). | en_US |
dc.Format.extent | 5.20 MB | en_US |
dc.language.iso | en_US | en_US |
dc.publisher | Springer-Verlag | en_US |
dc.subject | nAChR | en_US |
dc.subject | Functional characterization | en_US |
dc.subject | Planar lipid bilayers | en_US |
dc.subject | Integral membrane proteins | en_US |
dc.subject | Detergents | en_US |
dc.subject | Membrane lipids | en_US |
dc.subject | Biophysical characterization | en_US |
dc.title | Biophysical and Ion Channel Functional Characterization of the Torpedo Californica Nicotinic Acetylcholine Receptor in Varying Detergent-Lipid Environments | en_US |
dc.type | Article | en_US |
dc.rights.licence | Articles that are available through the PMC OAI and FTP services are still protected by copyright but are distributed under a Creative Commons or similar license that generally allows more liberal use than a traditional copyrighted work. Please refer to the license statement in each article for specific terms of use. The license terms are not identical for all the articles. | en_US |
dcterms.license | Articles that are available through the PMC OAI and FTP services are still protected by copyright but are distributed under a Creative Commons or similar license that generally allows more liberal use than a traditional copyrighted work. Please refer to the license statement in each article for specific terms of use. The license terms are not identical for all the articles. | en_US |
dcterms.rights | The respective copyright holders retain rights for reproduction, redistribution and reuse. | en_US |
dcterms.rightsHolder | The respective copyright holders retain rights for reproduction, redistribution and reuse. Users of PMC are directly and solely responsible for compliance with copyright restrictions and are expected to adhere to the terms and conditions defined by the copyright holder. | en_US |
dc.identifier.doi | DOI: 10.1007/s00232-008-9107-7 | en_US |
dc.local.Department | Department of Biology | en_US |
dc.local.Faculty | College of Natural Sciences | en_US |
dc.contributor.campus | University of Puerto Rico, Río Piedras Campus |
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