title: Characterization of dolichyl diphosphate phosphatase from rat liver.
abstract: Dolichyl diphosphate phosphatase (DolPPase) has been characterized in rat liver. Subcellular distribution studies indicate that the enzyme is localized in the endoplasmic reticulum. The in vitro enzymatic activity is stimulated by EDTA, due to release of inhibition by trivalent cations found in the assay tubes. All di- and trivalent cations tested were inhibitory, with the trivalent ions Al3+ and Fe3+ showing greater than 70% inhibition at a concentration of 10 microM. The assay requires the presence of a detergent for optimal activity, with Triton X-100 giving maximum activity at 0.1%. The substrate specificity of DolPPase toward polyprenyl diphosphates has been determined and indicates that there is little preference of the enzyme for substrates of different chain length, and either stereochemical orientation or degree of saturation of the alpha-isoprene unit. Km values of 11-14 microM were obtained for all substrates tested. Preliminary studies on the transmembrane topology of the DolPPase using latency assays, indicate that the active site of the enzyme may reside on the cytoplasmic face of the endoplasmic reticulum.
authors.affiliation: University of South Florida College of Medicine, Department of Biochemistry and Molecular Biology, Tampa 33612.
authors.forename: W L
authors.initials: WL
authors.lastname: Adair
authors.forename: N
authors.initials: N
authors.lastname: Cafmeyer
mesh_terms: D000818:Animals; D002412:Cations; D003902:Detergents / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000201:enzymology; D066298:In Vitro Techniques; D008099:Liver / Q000201:enzymology*; D008297:Male; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011755:Pyrophosphatases; D051381:Rats; D011919:Rats, Inbred Strains; D013347:Subcellular Fractions / Q000201:enzymology; D013379:Substrate Specificity
chemical_list: D002412:Cations; D003902:Detergents; D010744:Phosphoric Monoester Hydrolases; D011755:Pyrophosphatases; C023082:dolichyldiphosphatase