title: Retinyl ester hydrolases in retinal pigment epithelium.
abstract: In bovine retinal pigment epithelium membranes we have found three hydrolases which were active against trans-retinyl palmitate. This was possible by assaying different subcellular fractions as a function of pH in the range 3-9. Detection of these activities has been favored by the use in the enzyme assay of Triton X-100, which has an activating effect up to a concentration of 0.03% at a detergent-protein ratio of about 1.5-3.0. Apparent kinetic parameters for the retinyl ester hydrolases have been determined after a study of the optimization of assay conditions. Vmax values for hydrolases acting at pH 4.5, 6.0, and 7.0 were, respectively, 156, 55, and 70 nmol/h/mg. To identify the subcellular site for these hydrolytic activities, assays of marker enzymes from various organelles in each subcellular preparation were carried out, demonstrating the lysosomal origin of the pH 4.5 retinyl ester hydrolase and the microsomal origin of the pH 6.0 retinyl ester hydrolase and suggesting that the pH 7.0 retinyl ester hydrolase originates from the Golgi complex.
authors.affiliation: Istituto di Chimica Biologica, Facoltà di Medicina e Chirurgia, Università di Palermo, Italia.
authors.forename: M C
authors.initials: MC
authors.lastname: Gueli
authors.forename: C M
authors.initials: CM
authors.lastname: Nicotra
authors.forename: A M
authors.initials: AM
authors.lastname: Pintaudi
authors.forename: A
authors.initials: A
authors.lastname: Paganini
authors.forename: L
authors.initials: L
authors.lastname: Pandolfo
authors.forename: G
authors.initials: G
authors.lastname: De Leo
authors.forename: M A
authors.initials: MA
authors.lastname: Di Bella
mesh_terms: D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology; D002467:Cell Nucleus / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010857:Pigment Epithelium of Eye / Q000201:enzymology*; D013347:Subcellular Fractions / Q000201:enzymology
chemical_list: C022521:retinol-palmitate esterase; D002265:Carboxylic Ester Hydrolases