UIC-2012-082 – Luciferin-based biosensors to specifically detect peroxynitrite or hydrogen peroxide in vivo or in vitro

Hydrogen peroxide (H2O2) is a compound which is produced by living organisms to kill invading pathogens and for cellular signaling. Abnormal production of H2O2 results from oxidative stress related from aging, disease and injury. The study of H2O2 in relation to health of animals has been limited by a lack of methods to study H2O2 in the context of animal health and because of the transient nature of this molecule. Also deleterious to human health is peroxynitrite, a highly reactive biological oxidant formed under pathological conditions. This molecule has been implicated in a vast number of diseases from Alzheimer’s disease to atherosclerosis, stroke, respiratory diseases, inflammatory diseases, diabetes, pancreatitis, Huntington’s disease, and Parkinson’s disease. Peroxynitrite has the ability to move freely in cells through cell membranes and is known to nitrate tyrosine residues as well as to oxidize sulfhydryls, lipids, methionines and DNA. PCL-F is a novel boronate derivative of luciferin which can detect peroxynitrite and H2O2, both of which are indicators of oxidative stress. When PCL-F reacts with peroxynitrite or H2O2, the boronate moiety is released and allows the luciferin to be metabolized by luciferase. The resulting fluorescence can be measured from outside the skin to detect oxidative stress in a non-invasive manor in real time. PLC-1 is specific, sensitive, quantitative and versatile biosensor of peroxynitrite. The levels of oxidative stress due to hydrogen peroxide are determined in vivo by subtracting the level of oxidative stress detected by PLC-F (peroxynitrite and H2O2 signal) by PLC-1 (peroxynitrite signal). By using these two sensors the researcher or diagnostician can obtain real time non-invasive measurements of both hydrogen peroxide and peroxynitrite. Veronica Haywood vhaywo2@uic.otm.edu 312-996-4865