title: Detergent effects in rabbit liver microsomal UDP-glucuronosyltransferase studied by means of a continuous spectrophotometric assay with p-nitrophenol.
abstract: UDP-glucuronosyltransferase (UDPGT) has been studied with a continuous spectrophotometric assay employing UDP-glucuronic acid and p-nitrophenol as substrates. Activity is linearly dependent on the microsomal protein concentration. Male rabbit liver phenobarbital-induced microsomes exhibited a rate of 7.10 microM p-nitrophenol conjugated per minute at 37 degrees C. Addition of small amounts of Tergitol NP-10 caused an approximately 4-fold increase in conjugation activity; maximal activation was observed at 0.01% (v/v) detergent. However, inclusion of additional detergent caused significant inhibition of activity, such that 0.5% Tergitol caused the rate to fall 2.5-fold below the activity observed in the absence of detergent. Membrane solubilization was studied by light scattering. At maximal stimulation of p-nitrophenol UDPGT activity, the membrane was solubilized only approximately 17%. At the point of 50% solubilization, activity was still 91% of maximum. Complete membrane solubilization was achieved at approximately 0.2% Tergitol, and transferase activity had fallen slightly below the rate observed in the absence of detergent. Possible explanations for the unusual detergent-dependence of microsomal p-nitrophenol UDPGT activity are discussed.
authors.affiliation: Department of Biochemistry, University of Texas Health Center at Tyler 75710-2003.
authors.forename: S T
authors.initials: ST
authors.lastname: Martin
authors.forename: S D
authors.initials: SD
authors.lastname: Black
mesh_terms: D000818:Animals; D003902:Detergents; D014453:Glucuronosyltransferase / Q000032:analysis*; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D009596:Nitrophenols; D011060:Poloxalene; D011817:Rabbits; D012995:Solubility; D013053:Spectrophotometry / Q000379:methods*; D013379:Substrate Specificity
chemical_list: D003902:Detergents; D009596:Nitrophenols; D011060:Poloxalene; D014453:Glucuronosyltransferase; C024836:4-nitrophenol