The supramolecular structure of the GPCR rhodopsin in solution and native disc membranes.
Rhodopsin, the prototypical G-protein-coupled receptor, which is densely packed in the disc membranes of rod outer segments, was proposed to function as a monomer. However, a growing body of evidence indicates dimerization and oligomerization of numerous G-protein-coupled receptors, and atomic force microscopy images revealed rows of rhodopsin dimers in murine disc membranes. In this work we demonstrate by electron microscopy of negatively stained samples, blue native- and sodium dodecyl sulphate-polyacrylamide gel electrophoresis, chemical crosslinking, and by proteolysis that native bovine rhodopsin exists mainly as dimers and higher oligomers. These results corroborate the recent findings from atomic force microscopy and molecular modeling on the supramolecular structure and packing arrangement of murine rhodopsin dimers.
Suda
Kitaru
K
ME Muller Institute for Microscopy, Biozentrum, University of Basel, CH-4056 Basel, Switzerland.
Filipek
Slawomir
S
Palczewski
Krzysztof
K
Engel
Andreas
A
Fotiadis
Dimitrios
D
D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D003432:Cross-Linking Reagents / Q000737:chemistry; D019281:Dimerization; D046529:Microscopy, Electron, Transmission; D008958:Models, Molecular; D020836:Protein Structure, Quaternary; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D012243:Rhodopsin / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D012374:Rod Cell Outer Segment / Q000166:cytology* / Q000648:ultrastructure; D012995:Solubility; D012996:Solutions; D013388:Succinimides / Q000737:chemistry
D003432:Cross-Linking Reagents; D043562:Receptors, G-Protein-Coupled; D012996:Solutions; D013388:Succinimides; C085779:N-succinimidyl 6-(3-(2-pyridyldithio)propionamido)hexanoate; D012243:Rhodopsin; C011240:dithiobis(succinimidylpropionate)