title: Stabilization of acetylcholine receptor secondary structure by cholesterol and negatively charged phospholipids in membranes.
abstract: Fourier-transform infrared (FTIR) spectroscopy was used to study the secondary structure of purified Torpedo californica nicotinic acetylcholine receptor (AChR) in reconstituted membranes. Functional studies have previously demonstrated that the ion channel activity requires the presence of both sterol and negatively charged phospholipids in membranes. The present studies are designed to test the hypothesis that the alpha-helical structure of AChR may be stabilized by specific lipid molecules (sterol and/or negatively charged phospholipids) and that these alpha-helices may be responsible for the formation of a potential ion channel. FTIR data show statistically significant (p less than 0.005) spectral changes due to cholesterol and negatively charged phospholipids, respectively. On the basis of standard curves describing the relationship between the spectral properties and the secondary structural contents of water-soluble proteins, the observed spectral change at 931 cm-1 can be interpreted as an apparent change in the alpha-helix content from about 17% in the absence of sterols to about 20% in the presence of sterols, suggesting that protein-sterol interactions increase the helical structure of the AChR molecule. Similarly, the spectral change at 988 cm-1 can be interpreted as an apparent increase of beta-sheet content in the AChR molecule from about 20% to about 24% due to the presence of negatively charged phospholipids. Functional AChR in membranes thus appears to be correlated with higher alpha-helical and beta-sheet contents. It is concluded that one role of specific interactions between membrane protein and lipid molecules may be to maintain specific secondary structures necessary to support the ion channel function of AChR.
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000502:physiology*; D002784:Cholesterol / Q000494:pharmacology*; D005583:Fourier Analysis; D008563:Membrane Lipids / Q000378:metabolism; D010743:Phospholipids / Q000494:pharmacology*; D011487:Protein Conformation; D011950:Receptors, Cholinergic / Q000187:drug effects* / Q000302:isolation & purification; D013055:Spectrophotometry, Infrared / Q000379:methods; D013696:Temperature; D014101:Torpedo
chemical_list: D008563:Membrane Lipids; D010743:Phospholipids; D011950:Receptors, Cholinergic; D002784:Cholesterol