Assessment of the Mulder and Van Doorn kinetic procedure and rapid centrifugal analysis of UDP-glucuronosyltransferase activities.
The optimal experimental conditions of the enzyme assay described by Mulder and Van Doorn (1975, Biochem J. 151, 131-140) for the measurement of UDP-glucuronosyltransferase activities were tested towards structurally different aglycones. This assessment of this assay revealed that addition of Triton X-100 as enzyme activator was necessary because of its apparent inhibitory effects on interfering reactions. Under these conditions, accordance of the data with results published in the literature was obtained. We present for the first time an UDP-glucuronosyltransferase assay adapted on a fast analyser centrifuge which allows a rapid and sensitive measurement of enzyme activity that is very useful for kinetic constant determination, without consuming a large volume of reagents.
D000818:Animals; D002498:Centrifugation; D014453:Glucuronosyltransferase / Q000032:analysis*; D007700:Kinetics; D008297:Male; D008722:Methods; D008861:Microsomes / Q000201:enzymology; D009243:NAD; D017830:Octoxynol; D010636:Phenols; D011092:Polyethylene Glycols; D051381:Rats; D012997:Solvents; D013056:Spectrophotometry, Ultraviolet
D010636:Phenols; D012997:Solvents; D009243:NAD; D011092:Polyethylene Glycols; D017830:Octoxynol; D014453:Glucuronosyltransferase