Rod photoreceptors renew the membranous disks of the outer segments (ROS). New disks are assembled at the proximal base and old disks are shed at the distal tip. Rhodopsin, the major protein of the disk, remains with the disk into which it was inserted. Thus, it is true that the oldest rhodopsin is at the tip and the newest at the base. A microspectrophotometer is used to examine the properties...

Detergent-solubilized bovine rhodopsin produces mixed detergent/lipid/protein micelles. The effect of dodecyl maltoside detergent on the thermal stability of dark-state rhodopsin, and upon formation of the different intermediates after rhodopsin photobleaching (metarhodopsin II and metarhodopsin III), and upon transducin activation has been studied. No significant effect is observed for the the...

We compare the known retinitis pigmentosa (RP) mutations in rhodopsin with mutational data obtained for the complement factor 5a receptor (C5aR), a member of the rhodopsin-like family of G protein-coupled receptors (GPCRs). We have performed genetic analyses that define residues that are required for C5aR folding and function. The cognate residues in rhodopsin are not preferentially mutated in ...

We started a detailed characterization of adRP mutants in the transmembrane region of rhodopsin with a 2-fold interest, (i) elucidation of the molecular mechanism of the disease and (ii) structural and functional information, from the study of these mutations that might be relevant to other members of the GPCR superfamily. The rhodopsin crystal structure together with the available biochemical ...

The rhodopsin content of the human retina was determined spectrophotometrically. Retinas were removed completely by using an in vitro technique based on a simulation of retinal detachment. This method provides a total recovery of rod outer segments which contain the rhodopsin visual pigment or its apoprotein, opsin. Using this improved dissection method followed by a crude preparation of the re...

AIM To determine the pattern of rhodopsin mutations in Chinese retinitis pigmentosa (RP) patients. METHODS The rhodopsin gene was examined in 101 RP patients and 190 controls from Hong Kong. RESULTS Three coding changes were identified: Pro347Leu, Ala299Ser, and 5211delC. Each protein sequence alteration was found in one patient. Ala299Ser also existed in two controls. CONCLUSION The C-te...

Bathorhodopsin-rhodopsin difference spectra of native 11-cis-rhodopsin and regenerated 9-cis-rhodopsin were measured at room temperature with a double-beam laser spectrophotometer after excitation at 532 nm. A detailed analysis of data obtained at 85 psec after excitation suggests that the bathorhodopsins generated from 11-cis- and 9-cis-rhodopsin differ in their extinction coefficients and tha...

Introduction To investigate the conformational changes associated with photoactivation of rhodopsin [1], the GPCR of vision, we have embarked on a program of chemical synthesis of its transmembrane segments. By reconstituting such synthetic segments containing specific fluorescent and spin labels with expressed segments of rhodopsin, we hope to generate hybrid “split receptors” that are fully f...

A novel fluorescence method has been developed for detecting the light-induced conformational changes of rhodopsin and for monitoring the interaction between photolyzed rhodopsin and G-protein or arrestin. Rhodopsin in native membranes was selectively modified with fluorescent Alexa594-maleimide at the Cys(316) position, with a large excess of the reagent Cys(140) that was also derivatized. Mod...

PURPOSE To determine whether blue light induces photoreversal of rhodopsin bleaching in vivo. METHODS Eyes of anesthetized albino rats were exposed to either green (550 nm) or deep blue (403 nm) light, and the time course of rhodopsin bleaching was determined. Rhodopsin was isolated from whole retinas by detergent extraction and measured photometrically. To inhibit photoreversal of bleaching,...