Aryl hydrocarbon hydroxylase activity assayed in whole cell lysates using synchronous fluorescence spectroscopy.
Synchronous fluorescence spectrophotometry has been used to measure induced and constitutive levels of aryl hydrocarbon hydroxylase activity in lysates of C3H 10T1/2 mouse embryo fibroblasts. Without compromising sensitivity, the method was reproducible, eliminated the need to extract metabolites, and made the procedure simpler and less time consuming than other methods. Moreover, since the assay was tailored to directly measure 3-hydroxybenzo(a)pyrene, a metabolite produced by several cytochrome P-450s, it may be more generally applicable than dealkylation assays, which apparently detect only P-450-IA1.
Wells
R L
RL
Department of Radiological Health Sciences, Colorado State University, Fort Collins 80523.
Sneider
T W
TW
Elkind
M M
MM
D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000737:chemistry*; D001580:Benzopyrenes; D002457:Cell Extracts / Q000737:chemistry*; D002458:Cell Fractionation; D002460:Cell Line; D005347:Fibroblasts / Q000201:enzymology; D051379:Mice; D008809:Mice, Inbred C3H; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity
D001580:Benzopyrenes; D002457:Cell Extracts; C021338:3-hydroxybenzo(a)pyrene; D001189:Aryl Hydrocarbon Hydroxylases