Electron spin resonance studies of acyl chain motion in reconstituted nicotinic acetylcholine receptor membranes.
The electron spin resonance spectra of spin-label positional isomers of stearic acid (n-SASL) incorporated into nicotinic acetylcholine receptors (nAcChoR) reconstituted into dioleoylphosphatidylcholine (DOPC) were deconvoluted into bilayer- and protein-associated components by subtraction under conditions of slow exchange. The selectivity of n-SASL (n = 6, 9, 12, and 14) for the lipid-protein interface of the nAcChoR was threefold greater than that of DOPC and independent of the spin label position. The temperature at which exchange became apparent as judged from lineshape broadening of the mobile lipid component spectrum was dependent upon the position of the spin-label moiety; near the bilayer center, exchange broadening occurred at lower temperatures than it did closer to the lipid headgroup. This suggests that the lipid headgroup region of boundary lipids is relatively fixed, whereas its acyl chain whips on and off the protein with increasing frequency near the bilayer center. Motions on the microsecond time scale were examined by microwave power saturation. Each n-SASL saturated more readily when incorporated into vesicles containing the nAcChoR than when in pure DOPC liposomes. Therefore, lipid mobility is perturbed by the nAcChoR on the microsecond time scale with an apparent magnitude that is relatively modest, probably due to exchange on this time scale.
Department of Anesthesia, Massachusetts General Hospital, Boston 02114, USA.
D E
DE
Raines
G
G
Wu
L A
LA
Dalton
K W
KW
Miller
D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D004557:Electric Organ / Q000737:chemistry; D004578:Electron Spin Resonance Spectroscopy; D066298:In Vitro Techniques; D008081:Liposomes / Q000737:chemistry; D015394:Molecular Structure; D009038:Motion; D010713:Phosphatidylcholines / Q000737:chemistry; D011978:Receptors, Nicotinic / Q000737:chemistry*; D013113:Spin Labels; D013229:Stearic Acids / Q000737:chemistry; D013816:Thermodynamics; D014101:Torpedo
D008081:Liposomes; D010713:Phosphatidylcholines; D011978:Receptors, Nicotinic; D013113:Spin Labels; D013229:Stearic Acids; C017251:1,2-oleoylphosphatidylcholine