rutin and 3-4-methylenedioxy-beta-nitrostyrene

rutin has been researched along with 3-4-methylenedioxy-beta-nitrostyrene* in 1 studies

Other Studies

1 other study(ies) available for rutin and 3-4-methylenedioxy-beta-nitrostyrene

ArticleYear
Challenges in the evaluation of thiol-reactive inhibitors of human protein disulfide Isomerase.
    Free radical biology & medicine, 2017, Volume: 108

    This paper addresses how to evaluate the efficacy of the growing inventory of thiol-reactive inhibitors of mammalian protein disulfide Isomerase (PDI) enzymes under realistic concentrations of potentially competing thiol-containing peptides and proteins. For this purpose, we introduce a variant of the widely-used reductase assay by using a commercially-available cysteine derivative (BODIPY FL L-Cystine; BD-SS) that yields a 55-fold increase in fluorescence (excitation/emission; 490/513nm) on scission of the disulfide bond. This plate reader-compatible method detects human PDI down to 5-10nM, can utilize a range of thiol substrates (including 5µM dithiothreitol, 10µM reduced RNase thiols, and 5mM glutathione; GSH), and can operate from pH 6-9.5 in a variety of buffers. PDI assays often employ low micromolar levels of substrates leading to ambiguities when thiol-directed inhibitors are evaluated. The present work utilizes 5mM GSH for both pre-incubation and assay phases to more realistically reflect the high concentration of thiols that an inhibitor would encounter intracellularly. Extracellular PDI faces a much lower concentration of potentially competing thiols; to assess reductase activity under these conditions, the pre-reduced PDI is treated with inhibitor and then fluorescence increase upon reduction of BD-SS is followed in the absence of additional competing thiols. Both assay modes were tested with four mechanistically diverse PDI inhibitors. Two reversible reagents, 3,4-methylenedioxy-β-nitrostyrene (MNS) and the arsenical APAO, were found to be strong inhibitors of PDI in the absence of competing thiols, but were ineffective in the presence of 5mM GSH. A further examination of the nitrostyrene showed that MNS not only forms facile Michael adducts with GSH, but also with the thiols of unfolded proteins (K

    Topics: Animals; Arsenicals; Binding Sites; Boron Compounds; Cell-Free System; Cysteine; Cystine; Dioxolanes; Glucosides; Glutathione; Humans; Mammals; Oxidation-Reduction; Protein Disulfide-Isomerases; Quercetin; Spectrometry, Fluorescence; Sulfhydryl Compounds

2017