title: A high-throughput turbidometric assay for screening inhibitors of Leishmania major protein disulfide isomerase.
abstract: The use of a high-throughput technique to perform a pilot screen for Leishmania major protein disulfide isomerase (LmPDI) inhibitors identification is reported. In eukaryotic cells, protein disulfide isomerase (PDI) plays a crucial role in protein folding by catalyzing the rearrangement of disulfide bonds in substrate proteins following their synthesis. LmPDI displays similar domain structure organization and functional properties to other PDI family members and is involved in Leishmania virulence. The authors used a method based on the enzyme-catalyzed reduction of insulin in the presence of dithiothreitol. The screen of a small library of 1920 compounds was performed in a 384-well format and led to the identification of 27 compounds with inhibitory activity against LmPDI. The authors further tested the cytotoxicity of these compounds using Jurkat cells as well as their effect on Leishmania donovani amastigotes using high-content analysis. Results show hexachlorophene and a mixture of theaflavin monogallates inhibit Leishmania multiplication in infected macrophages derived from THP-1 cells, although the inhibitory effect on LmPDI enzymatic activity does not necessarily correlate with the antileishmanial activity.
authors: Laboratory of Immunopathology Vaccinology and Molecular Genetics (LIVGM), Institut Pasteur de Tunis, Tunis-Belvédère, Tunisia.
authors: Noureddine
authors: N
authors: Ben Khalaf
authors: Géraldine
authors: G
authors: De Muylder
authors: Joseline
authors: J
authors: Ratnam
authors: Kenny
authors: K
authors: Kean-Hooi Ang
authors: Michelle
authors: M
authors: Arkin
authors: James
authors: J
authors: McKerrow
authors: Mehdi
authors: M
authors: Chenik
mesh_terms: D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D055808:Drug Discovery / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D057166:High-Throughput Screening Assays; D006801:Humans; D019169:Jurkat Cells; D007893:Leishmania donovani / Q000187:drug effects; D018320:Leishmania major / Q000201:enzymology*; D019704:Protein Disulfide-Isomerases / Q000037:antagonists & inhibitors*; D054852:Small Molecule Libraries / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; D054852:Small Molecule Libraries; D019704:Protein Disulfide-Isomerases