title: On the use of peroxy-caged luciferin (PCL-1) probe for bioluminescent detection of inflammatory oxidants in vitro and in vivo - Identification of reaction intermediates and oxidant-specific minor products.
abstract: Peroxy-caged luciferin (PCL-1) probe was first used to image hydrogen peroxide in living systems (Van de Bittner et al., 2010 [9]). Recently this probe was shown to react with peroxynitrite more potently than with hydrogen peroxide (Sieracki et al., 2013 [11]) and was suggested to be a more suitable probe for detecting peroxynitrite under in vivo conditions. In this work, we investigated in detail the products formed from the reaction between PCL-1 and hydrogen peroxide, hypochlorite, and peroxynitrite. HPLC analysis showed that hydrogen peroxide reacts slowly with PCL-1, forming luciferin as the only product. Hypochlorite reaction with PCL-1 yielded significantly less luciferin, as hypochlorite oxidized luciferin to form a chlorinated luciferin. Reaction between PCL-1 and peroxynitrite consists of a major and minor pathway. The major pathway results in luciferin and the minor pathway produces a radical-mediated nitrated luciferin. Radical intermediate was characterized by spin trapping. We conclude that monitoring of chlorinated and nitrated products in addition to bioluminescence in vivo will help identify the nature of oxidant responsible for bioluminescence derived from PCL-1.
authors.affiliation: Department of Biophysics and Free Radical Research Center, Medical College of Wisconsin, Milwaukee, WI 53226, United States. Electronic address: jzielonk@mcw.edu.
authors.forename: Jacek
authors.initials: J
authors.lastname: Zielonka
authors.affiliation: Institute of Polymer and Dye Technology, Faculty of Chemistry, Lodz University of Technology, Stefanowskiego 12/16, 90-924 Lodz, Poland. Electronic address: radoslaw.podsiadly@p.lodz.pl.
authors.forename: Radosław
authors.initials: R
authors.lastname: Podsiadły
authors.affiliation: Department of Biophysics and Free Radical Research Center, Medical College of Wisconsin, Milwaukee, WI 53226, United States. Electronic address: mzielonka@mcw.edu.
authors.forename: Monika
authors.initials: M
authors.lastname: Zielonka
authors.affiliation: Aix-Marseille Université, CNRS, ICR UMR 7273, 13397 Marseille, France. Electronic address: micael.hardy@univ-provence.fr.
authors.forename: Micael
authors.initials: M
authors.lastname: Hardy
authors.affiliation: Department of Biophysics and Free Radical Research Center, Medical College of Wisconsin, Milwaukee, WI 53226, United States. Electronic address: balarama@mcw.edu.
authors.forename: Balaraman
authors.initials: B
authors.lastname: Kalyanaraman
mesh_terms: D000818:Animals; D002460:Cell Line; D049454:Firefly Luciferin; D006861:Hydrogen Peroxide / Q000032:analysis*; D006997:Hypochlorous Acid / Q000032:analysis*; D007249:Inflammation; D008163:Luminescent Measurements / Q000379:methods*; D008264:Macrophages / Q000378:metabolism* / Q000648:ultrastructure; D051379:Mice; D015335:Molecular Probes / Q000138:chemical synthesis / Q000737:chemistry*; D010084:Oxidation-Reduction; D030421:Peroxynitrous Acid / Q000032:analysis*; D019097:Spin Trapping
chemical_list: D015335:Molecular Probes; D030421:Peroxynitrous Acid; D049454:Firefly Luciferin; D006997:Hypochlorous Acid; D006861:Hydrogen Peroxide
keywords: Bioluminescence; Boronate probes; Luciferin; Spin traps