Identification and immunohistochemistry of retinol dehydrogenase from bovine retinal pigment epithelium.
We studied the properties of retinol dehydrogenase (11-cis-specific) from bovine retinal pigment epithelium. Detergents caused a loss of retinol dehydrogenase activity; therefore, we added 3 mM NADH as a stabilizer to solubilize this enzyme and partially purified this enzyme using Sepharose CL-6B and hydroxyapatite column chromatography. The partially-purified sample, which contained two major proteins (66 kDa, 33 kDa), had substrate preference to 11-cis and 13-cis-retinal but not to all-trans and 9-cis isomers. Monoclonal anti-33 kDa protein of retinal pigment epithelial crude extract by Western blotting. In addition, we found that monoclonal anti-retinol dehydrogenase antibody bound specifically to retinal pigment epithelium and not to Müller cells or to rod outer segments by immunohistochemical methods.
Suzuki
Y
Y
Department of Ophthalmology, Tohoku University School of Medicine, Sendai, Japan.
Ishiguro
S
S
Tamai
M
M
D000429:Alcohol Oxidoreductases / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D000818:Animals; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D002417:Cattle; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007150:Immunohistochemistry; D008861:Microsomes / Q000201:enzymology; D010857:Pigment Epithelium of Eye / Q000201:enzymology*; D012160:Retina / Q000201:enzymology*; D012172:Retinaldehyde / Q000378:metabolism; D013237:Stereoisomerism; D014801:Vitamin A / Q000378:metabolism
D000906:Antibodies; D014801:Vitamin A; D000429:Alcohol Oxidoreductases; C022520:retinol dehydrogenase; D012172:Retinaldehyde