Conformational states and dynamics of rhodopsin in micelles and bilayers.
Nitroxide sensors were placed in rhodopsin at sites 140, 227, 250, and 316 to monitor the dynamics and conformation of the receptor at the cytoplasmic surface in solutions of dodecyl maltoside (DM), digitonin, and phospholipid bilayers of two compositions. The EPR spectra reveal a remarkable similarity of rhodopsin structure and the activating conformational change in DM and bilayers, the hallmark of which is an outward tilt of transmembrane helix VI. This conformational change is blocked in solutions of digitonin, although changes in optical absorbance accompany activation, showing that absorbance and structural changes are not necessarily coupled. In DM and bilayers, the receptor is apparently in equilibrium between conformational substates whose populations are modulated by activation. Despite the general similarity in the two environments, the receptor conformations have increased flexibility in DM relative to bilayers. For the activated receptor in DM and bilayers, a pH-dependent conformational equilibrium is identified that may correspond to the optically characterized MII(a)()-MII(b)() equilibrium. No specific effects of headgroup composition on receptor conformation in lipid bilayers were found.
Kusnetzow
Ana Karin
AK
Jules Stein Eye Institute and Department of Chemistry and Biochemistry, University of California, Los Angeles, California 90095-7008, USA.
Altenbach
Christian
C
Hubbell
Wayne L
WL
D019943:Amino Acid Substitution; D000818:Animals; D003497:Cyclic N-Oxides / Q000737:chemistry; D004072:Digitonin / Q000737:chemistry; D004578:Electron Spin Resonance Spectroscopy; D005960:Glucosides / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry; D008567:Membranes, Artificial; D008698:Mesylates / Q000737:chemistry; D008823:Micelles; D011487:Protein Conformation; D012243:Rhodopsin / Q000737:chemistry* / Q000235:genetics; D013113:Spin Labels
D003497:Cyclic N-Oxides; D005960:Glucosides; D008051:Lipid Bilayers; D008567:Membranes, Artificial; D008698:Mesylates; D008823:Micelles; D013113:Spin Labels; C045880:methanesulfonic acid; C040358:dodecyl maltoside; D012243:Rhodopsin; D004072:Digitonin