Average membrane penetration depth of tryptophan residues of the nicotinic acetylcholine receptor by the parallax method.
The membrane penetration depths of tryptophan residues in the nicotinic acetylcholine receptor from Torpedo californica have been analyzed in reconstituted membranes containing purified receptor and defined lipids. Dioleoylphosphatidylcholine and three spin-labeled phosphatidylcholines with the nitroxide group at three different positions on the fatty acyl chain were used for reconstitution of the receptor. The spin-labeled phospholipids serve as quenchers of tryptophan fluorescence. Differential quenching of the intrinsic fluorescence of the acetylcholine receptor by the spin-labeled phospholipids has been utilized to analyze the average membrane penetration depth of tryptophans by the parallax method [Chattopadhyay, A., & London, E. (1987) Biochemistry 26, 39-45]. Analyses of the quenching data indicate that the tryptophan residues on the average are at a shallow location (10.1 A from the center of the bilayer) in the membrane. In addition, the generally low levels of quenching imply that the majority of tryptophan residues are located in the putative extramembranous region of the receptor. These results are consistent with several proposed models for the tertiary structure of the acetylcholine receptor and are relevant to ongoing analyses of the overall conformation and orientation of the acetylcholine receptor in the membrane.
Chattopadhyay
A
A
Department of Biochemistry and Biophysics, University of California, Davis 95616.
McNamee
M G
MG
D000109:Acetylcholine / Q000378:metabolism*; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D005454:Fluorescence Polarization; D010713:Phosphatidylcholines / Q000378:metabolism*; D010743:Phospholipids; D011978:Receptors, Nicotinic / Q000378:metabolism*; D014101:Torpedo; D014364:Tryptophan / Q000378:metabolism*
D010713:Phosphatidylcholines; D010743:Phospholipids; D011978:Receptors, Nicotinic; D014364:Tryptophan; C017251:1,2-oleoylphosphatidylcholine; D000109:Acetylcholine