NDLI: Assessing the Lipid Requirements of the Torpedo californica Nicotinic Acetylcholine Receptor†

Content Provider	PubMed Central
Author	Hamouda, Ayman K. Sanghvi, Mitesh Sauls, Daniel Machu, Tina K. Blanton, Michael P.
Abstract	The lipid requirements of the Torpedo californica nicotinic acetylcholine receptor (nAChR) were assessed by reconstituting purified receptors into lipid vesicles of defined composition and by using photolabeling with 3-trifluoromethyl-3-(m-[125I]iodophenyl) diazirine ([125I]TID) to determine functionality. Earlier studies demonstrated that nAChRs reconstituted into membranes containing phosphatidylcholine (PC), the anionic lipid phosphatidic acid (PA) and cholesterol (CH) are particularly effective at stabilizing the nAChR in the resting (closed) state that is capable of undergoing agonist-induced conformational transitions (i.e. functionality). The present studies demonstrate that: (1) there is no obligatory requirement for PC; (2) that increasing the CH content serves to increase the degree to which nAChRs are stabilized in the resting state, this effect saturates at ∼ 35 mol% (molar lipid percentage); (3) the effect of increasing levels of PA saturates at ∼12 mol% and in the absence of PA nAChRs are stabilized in the desensitized state (i.e. nonfunctional). Native Torpedo membranes contain ∼35 mol% CH but less than 1 mol% PA, suggesting that other anionic lipid may substitute for PA. We report that: (4) phosphatidylserine (PS) and phosphatidylinositol (PI), anionic lipids that are abundant in native Torpedo membranes, also stabilize the receptor in the resting state although with reduced efficacy (∼50-60%) compared to PA; (5) For nAChRs reconstituted into PA/CH membranes at different lipid-protein molar ratios, receptor functionality decreases rapidly below ∼65 lipids per receptor. Collectively, these results are consistent with a functional requirement of a single shell of lipids surrounding the nAChR and specific anionic lipid- and sterol- (CH) protein interactions.
Related Links	http://dx.doi.org/10.1021/bi052281z
Ending Page	4337
Page Count	11
Starting Page	4327
File Format	PDF
ISSN	00062960
e-ISSN	15204995
Journal	Biochemistry
Issue Number	13
Volume Number	45
Language	English
Publisher Date	2006-04-01
Access Restriction	Open
Subject Keyword	Biochemistry Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Biochemistry

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Assessing the Lipid Requirements of the Torpedo californica Nicotinic Acetylcholine Receptor†

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