redox-based inactivation of cysteine cathepsins by compounds containing the 4-aminophenol moietyredox-based半胱氨酸组织蛋白酶的失活化合物包含4-aminophenol一半.pdfVIP

Redox-Based Inactivation of Cysteine Cathepsins by Compounds Containing the 4-Aminophenol Moiety ´ 1 ˇ 2 2 1,3 Bojana Mirkovic *, Izidor Sosic , Stanislav Gobec , Janko Kos 1 Department of Pharmaceutical Biology, Faculty of Pharmacy, University of Ljubljana, Ljubljana, Slovenia, 2 Department of Medicinal Chemistry, Faculty of Pharmacy, ˇ University of Ljubljana, Ljubljana, Slovenia, 3 Department of Biotechnology, Jozef Stefan Institute, Ljubljana, Slovenia Abstract Background: Redox cycling compounds have been reported to cause false positive inhibition of proteases in drug discovery studies. This kind of false positives can lead to unusually high hit rates in high-throughput screening campaigns and require further analysis to distinguish true from false positive hits. Such follow-up studies are both time and resource consuming. Methods and Findings: In this study we show that 5-aminoquinoline-8-ol is a time-dependent inactivator of cathepsin B with a kinact/KI of 36.7 613.6 M21s21 using enzyme kinetics. 5-Aminoquinoline-8-ol inhibited cathepsins H, L and B in the same concentration range, implying a non-specific mechanism of inhibition. Further analogues, 4-aminonaphthalene-1-ol and 4-aminophenol, also displayed time-dependent inhibition of cathepsin B with kinact/KI values of 406.4 610.8 and 36.561.3 M21s21. No inactivation occurred in the absence of either the amino or the hydroxyl group, suggesting that the 4- aminophenol moiety is a prerequisite for enzyme inactivation. Induction of redox oxygen species (ROS) by 4-aminophenols in various redox environments was determined by the fluorescent probe 29,7 9-dichl