Cell Biochemistry and Biophysics

, Volume 75, Issue 3–4, pp 335–349 | Cite as

Recent Developments in the Probes and Assays for Measurement of the Activity of NADPH Oxidases

  • Jacek Zielonka
  • Micael Hardy
  • Radosław Michalski
  • Adam Sikora
  • Monika Zielonka
  • Gang Cheng
  • Olivier Ouari
  • Radosław Podsiadły
  • Balaraman Kalyanaraman
Original Paper

Abstract

NADPH oxidases are a family of enzymes capable of transferring electrons from NADPH to molecular oxygen. A major function of NADPH oxidases is the activation of molecular oxygen into reactive oxygen species. Increased activity of NADPH oxidases has been implicated in various pathologies, including cardiovascular disease, neurological dysfunction, and cancer. Thus, NADPH oxidases have been identified as a viable target for the development of novel therapeutics exhibiting inhibitory effects on NADPH oxidases. Here, we describe the development of new assays for measuring the activity of NADPH oxidases enabling the high-throughput screening for NADPH oxidase inhibitors.

Keywords

NADPH oxidase Reactive oxygen species Fluorescent probes High-throughput screening HPLC 

Abbreviations

2-OH-E+

2-hydroxyethidium

2-OH-Pr2+

2-hydroxypropidium

BMPO

5-tert-butoxycarbonyl-5-methyl-1-pyrroline-N-oxide

CAT

catalase

CBA

coumarin-7-boronic acod

CD

cyclodextrin

COH

7-hydroxycoumarin

cyt c3+

ferricytochrome c

DEPMPO

5-(diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide

DIPPMPO

5-(diisopropoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide

DMPO

5,5-dimethyl-1-pyrroline-N-oxide

DPI

diphenyleneiodonium

E+

ethidium

E+-E+

diethidium

EPR

electron paramagnetic resonance

GSH

glutathione

HE

hydroethidine (or dihydroethidium)

HE•+

HE radical cation

HPLC

high-performance liquid chromatography

HPr+

hydropropidine

HRP

horseradish peroxidase

HTS

high-throughput screening

L-012

8-amino-5-chloro-2,3-dihydro-7-phenyl-pyrido[3,4-d]pyridazine

MPO

myeloperoxidase

NADPH

nicotinamide adenine dinucleotide phosphate, reduced form

NBT

nitroblue tetrazolium

Nox2

NADPH oxidase-2

Nox4

NADPH oxidase-4

Nox5

NADPH oxidase-5

OCR

oxygen consumption rate

PMA

phorbol 12-myristate 13-acetate

SOD

superoxide dismutase

VAS2870

1,3-benzoxazol-2-yl-3-benzyl-3H-[1–3]triazolo[4,5-d]pyrimidin-7-yl sulfide

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Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  1. 1.Department of Biophysics and Free Radical Research CenterMedical College of WisconsinMilwaukeeUSA
  2. 2.Aix Marseille Univ, CNRS, ICRMarseilleFrance
  3. 3.Institute of Applied Radiation Chemistry, Faculty of ChemistryLodz University of TechnologyLodzPoland
  4. 4.Institute of Polymer and Dye Technology, Faculty of ChemistryLodz University of TechnologyLodzPoland

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