Isomerization of all-trans-9- and 13-desmethylretinol by retinal pigment epithelial cells.
Photoisomerization of 11-cis-retinal to all-trans-retinal triggers phototransduction in the retinal photoreceptor cells and causes ultimately the sensation of vision. 11-cis-Retinal is enzymatically regenerated through a complex set of reactions in adjacent retinal pigment epithelial cells (RPE). In this study using all-trans-9-desmethylretinol (lacking the C(19) methyl group) and all-trans-13-desmethylretinol (lacking the C(20) methyl group), we explored the effects of C(19) and C(20) methyl group removals on isomerization of these retinols in RPE microsomes. The C(19) methyl group may be involved in the substrate activation, whereas the C(20) methyl group causes steric hindrance with a proton in position C(10) of 11-cis-retinol; thus, removal of this group could accelerate isomerization. We found that all-trans-9-desmethylretinol and all-trans-13-desmethylretinol are isomerized to their corresponding 11-cis-alcohols, although with lower efficiencies than isomerization of all-trans-retinol to 11-cis-retinol. These findings make the mechanism of isomerization through the C(19) methyl group unlikely, because in the case of 9-desmethylretinol, the isomerization would have to progress by proton abstraction from electron-rich olefinic C(9). The differences between all-trans-retinol, all-trans-9-desmethylretinol, and all-trans-13-desmethylretinol appear to be a consequence of the enzymatic properties, and binding affinities of the isomerization system, rather than differences in the chemical or thermodynamic properties of these compounds. This observation is also supported by quantum chemical calculations. It appears that both methyl groups are not essential for the isomerization reaction and are not likely involved in formation of a transition stage during the isomerization process.
Stecher
H
H
Department of Ophthalmology, University of Washington, Seattle 98195, USA.
Prezhdo
O
O
Das
J
J
Crouch
R K
RK
Palczewski
K
K
D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001059:Apoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002417:Cattle; D004224:Diterpenes; D007536:Isomerism; D010857:Pigment Epithelium of Eye / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D011487:Protein Conformation; D011789:Quantum Theory; D012172:Retinaldehyde / Q000737:chemistry* / Q000378:metabolism; D012176:Retinoids / Q000737:chemistry / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000502:physiology
C020932:11-cis-retinal-binding protein; D001059:Apoproteins; D002352:Carrier Proteins; D004224:Diterpenes; D012176:Retinoids; D012710:Serum Albumin, Bovine; C031338:13-cis-retinal; C031390:9-cis-retinal; D000429:Alcohol Oxidoreductases; C022520:retinol dehydrogenase; D012172:Retinaldehyde