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. Free Radic Biol Med 2016 Oct;99:32-42
Date
10/23/2016Pubmed ID
27458121Pubmed Central ID
PMC5107150DOI
10.1016/j.freeradbiomed.2016.07.023Scopus ID
2-s2.0-84979887634 (requires institutional sign-in at Scopus site) 45 CitationsAbstract
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.
Author List
Zielonka J, Podsiadły R, Zielonka M, Hardy M, Kalyanaraman BAuthors
Micael Joel Hardy PhD Visiting Assistant Professor in the Biophysics department at Medical College of WisconsinBalaraman Kalyanaraman PhD Professor in the Biophysics department at Medical College of Wisconsin
Jacek M. Zielonka PhD Assistant Professor in the Biophysics department at Medical College of Wisconsin
MESH terms used to index this publication - Major topics in bold
AnimalsCell Line
Firefly Luciferin
Hydrogen Peroxide
Hypochlorous Acid
Inflammation
Luminescent Measurements
Macrophages
Mice
Molecular Probes
Oxidation-Reduction
Peroxynitrous Acid
Spin Trapping
