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Sci Rep. 2016 Sep 19;6:32766. doi: 10.1038/srep32766.

Uncovering the lipidic basis for the preparation of functional nicotinic acetylcholine receptor detergent complexes for structural studies.

Author information

1
Department of Physical Sciences, University of Puerto Rico, Río Piedras Campus, PO Box 23323, San Juan, 00931-03323, Puerto Rico.
2
Department of Chemistry, University of Puerto Rico, Río Piedras Campus, PO Box 23346, San Juan, 00931-3346, Puerto Rico.
3
Department of Biology, University of Puerto Rico, Río Piedras Campus, PO Box 70377, San Juan, 00936-8377, Puerto Rico.
4
Department of Pharmaceutical Sciences, School of Pharmacy, University of Puerto Rico, San Juan, 00936, Puerto Rico.
5
University of Puerto Rico, Molecular Sciences and Research Center, San Juan, 00926, Puerto Rico.

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.

PMID:
27641515
PMCID:
PMC5027579
DOI:
10.1038/srep32766
[Indexed for MEDLINE]
Free PMC Article

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