Response to Pick JAQUET , Vincent , RUTTER , A

In his letter, Dr. Pick criticizes our use of relative values when representing the NOX2 inhibitory action of a novel small molecule (GSK2795039) in a semi-recombinant NOX2 membrane assay. To address this concern, we performed additional experiments using the superoxide inhibitable assays cytochrome C and water soluble tetrazolium salt (WST-1) reduction. In this letter, we document turnover values between 80 and 100 mol O2(•-)/s/mol cytochrome b558 in our semi-recombinant assay and confirmed that GSK2795039 inhibits the NOX2 isoform in the submicromolar range. Antioxid. Redox Signal. 23, 1251-1253. JAQUET, Vincent, RUTTER, A Richard. Response to Pick. Antioxidants & Redox Signaling, 2015, vol. 23, no. 15, p. 1251-1253 DOI : 10.1089/ars.2015.6565

[1]  C. Davies,et al.  Discovery of GSK2795039, a Novel Small Molecule NADPH Oxidase 2 Inhibitor. , 2015, Antioxidants & redox signaling.

[2]  FlemingIngrid,et al.  Unchanged NADPH Oxidase Activity in Nox1-Nox2-Nox4 Triple Knockout Mice: What Do NADPH-Stimulated Chemiluminescence Assays Really Detect? , 2015 .

[3]  Y. Berdichevsky,et al.  The dehydrogenase region of the NADPH oxidase component Nox2 acts as a protein disulfide isomerase (PDI) resembling PDIA3 with a role in the binding of the activator protein p67phox , 2015, Front. Chem..

[4]  E. Pick.  Cell-free NADPH oxidase activation assays: "in vitro veritas". , 2014, Methods in molecular biology.

[5]  K. Krause,et al.  Detection of reactive oxygen species derived from the family of NOX NADPH oxidases. , 2012, Free radical biology & medicine.

[6]  D. Grunwald,et al.  Role of Putative Second Transmembrane Region of Nox2 Protein in the Structural Stability and Electron Transfer of the Phagocytic NADPH Oxidase* , 2011, The Journal of Biological Chemistry.

[7]  D. Grunwald,et al.  Regulation of NADPH Oxidase Activity in Phagocytes , 2010, The Journal of Biological Chemistry.

[8]  T. Kawahara,et al.  Constitutive NADPH-Dependent Electron Transferase Activity of the Nox4 Dehydrogenase Domain† , 2010, Biochemistry.

[9]  M. Berridge,et al.  Superoxide produced by activated neutrophils efficiently reduces the tetrazolium salt, WST-1 to produce a soluble formazan: a simple colorimetric assay for measuring respiratory burst activation and for screening anti-inflammatory agents. , 2000, Journal of immunological methods.