Calorimetric studies of bovine rod outer segment disk membranes support a monomeric unit for both rhodopsin and opsin.
The photoreceptor rhodopsin is a G-protein coupled receptor that has recently been proposed to exist as a dimer or higher order oligomer, in contrast to the previously described monomer, in retinal rod outer segment disk membranes. Rhodopsin exhibits considerably greater thermal stability than opsin (the bleached form of the receptor), which is reflected in an approximately 15 degrees C difference in the thermal denaturation temperatures (T(m)) of rhodopsin and opsin as measured by differential scanning calorimetry. Here we use differential scanning calorimetry to investigate the effect of partial bleaching of disk membranes on the T(m) of rhodopsin and of opsin in native disk membranes, as well as in cross-linked disk membranes in which rhodopsin dimers are known to be present. The T(m)s of rhodopsin and opsin are expected to be perturbed if mixed oligomers are present. The T(m) remained constant for rhodopsin and opsin in native disks regardless of the level of bleaching. In contrast, the T(m) of cross-linked rhodopsin in disk membranes was dependent on the extent of bleaching. The energy of activation for denaturation of rhodopsin and cross-linked rhodopsin was calculated. Cross-linking rhodopsin significantly decreased the energy of activation. We conclude that in native disk membranes, rhodopsin behaves predominantly as a monomer.
Edrington
Thomas C
TC
Department of Molecular and Cell Biology, University of Connecticut, Storrs, Connecticut 06250, USA.
Bennett
Michael
M
Albert
Arlene D
AD
D000818:Animals; D002152:Calorimetry, Differential Scanning; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism*; D003432:Cross-Linking Reagents / Q000378:metabolism; D019281:Dimerization; D007700:Kinetics; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary; D012243:Rhodopsin / Q000737:chemistry / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000166:cytology*; D017299:Rod Opsins / Q000737:chemistry / Q000378:metabolism*; D044366:Transition Temperature
D003432:Cross-Linking Reagents; D017299:Rod Opsins; D012243:Rhodopsin