title: Detection of Total UDP-Glucuronosyltransferase (UGT) Activity in Melanoma Cells.
abstract: The UDP-glucuronosyltransferases (UGTs) are integrally involved in the clearance of a wide range of drugs used to combat human diseases. UGT expression levels and activity can be induced by drug addition to cells and has been proposed as a potential intratumoral drug resistance mechanism. Traditional methods of assaying UGT activity are drug-centric and require HPLCs with multiple detectors (dependent on individual drug). Here, we describe a generalized method to detect total UGT activity (intrinsic or induced) via the UGT-Glo assay which utilizes a general UGT substrate with luminescence as the readout eliminating the need for multiple HPLC detectors to detect total UGT activity in a given sample. The method detailed here can be applied for any UGT containing sample, allowing for the efficient detection of total UGT activity to be the functional endpoint using a plate reader. In this manner, global changes in UGT activity can be monitored in response to a wide variety of stimuli.
authors.affiliation: Chao Family Comprehensive Cancer Center and Departments of Medicine (RWD, FLM), Biological Chemistry (FLM), Epidemiology (RWD) and Public Health (FLM), University of California Irvine, Sprague Hall B200, Irvine, CA, 92697, USA. rdelling@uci.edu.
authors.forename: Ryan W
authors.initials: RW
authors.lastname: Dellinger
authors.affiliation: Chao Family Comprehensive Cancer Center and Departments of Medicine (RWD, FLM), Biological Chemistry (FLM), Epidemiology (RWD) and Public Health (FLM), University of California Irvine, Sprague Hall B200, Irvine, CA, 92697, USA.
authors.forename: Frank L
authors.initials: FL
authors.lastname: Meyskens
keywords: Drug resistance; Glucuronidase activity; Melanoma; UDP-glucuronosyltransferase; UGT-Glo