Peptide‐Functionalized Fluorescent Particles for In Situ Detection of Nitric Oxide via Peroxynitrite‐Mediated Nitration

Abstract

Nitric oxide (NO) is a free radical signaling molecule that plays a crucial role in modulating physiological homeostasis across multiple biological systems. NO dysregulation is linked to the pathogenesis of multiple diseases; therefore, its quantification is important for understanding pathophysiological processes. The detection of NO is challenging, typically limited by its reactive nature and short half‐life. Additionally, the presence of interfering analytes and accessibility to biological fluids in the native tissues make the measurement technically challenging and often unreliable. Here, a bio‐inspired peptide‐based NO sensor is developed, which detects NO‐derived oxidants, predominately peroxynitrite‐mediated nitration of tyrosine residues. It is demonstrated that these peptide‐based NO sensors can detect peroxynitrite‐mediated nitration in response to physiological shear stress by endothelial cells in vitro. Using the peptide‐conjugated fluorescent particle immunoassay, peroxynitrite‐mediated nitration activity with a detection limit of ≈100 × 10−9 m is detected. This study envisions that the NO detection platform can be applied to a multitude of applications including monitoring of NO activity in healthy and diseased tissues, localized detection of NO production of specific cells, and cell‐based/therapeutic screening of peroxynitrite levels to monitor pronitroxidative stress in biological samples.

Progress and Promise of Nitric Oxide‐Releasing Platforms

“…Chang et al recently reported a peptide-based NO sensor that can resolve nanomolar concentrations of NO while providing information within biological systems (detecting NO in response to physiological levels of shear stress). [133] This sensor may be introduced intravenously to assess NO levels in the circulation or at specific tissues. This example demonstrates the type of sensors that stand to make impact for the development of personalized NO therapies.…”

Construction of a Rational-Designed Multifunctional Platform Based on a Fluorescence Resonance Energy Transfer Process for Simultaneous Detection of pH and Endogenous Peroxynitrite

“…In 2006, Yang reported the first ONOO – specific-recognition probe based on fluorescein derivatives and applied it to cell imaging . Since then, many fluorescent probes based on chromophores such as fluorescein, rhodamine derivatives, − coumarin, naphthylamides, and cyanine analogues have been reported for ONOO – recognition and applied in live organisms. , These fluorescent probes exhibit high sensitivity and selectivity for ONOO – measurement; however, most of the above studies only focused on ONOO – detection; other factors that may affect the stability and reactivity of ONOO – were given little consideration, such as solvents, ions, and pH of the biological environment.…”

Ultra-Rapid Detection of Endogenous Nitric Oxide Based on Fluorescent Conjugated Polymers Probe

“…The fluorescence technique is advantageous in nondestructive testing. Many fluorescent probes have been developed to detect endogenous NO. − Especially, o-diaminophenyl group based fluorescence techniques are very popular for the detection of NO owing to their sensitivity, selectivity, simplicity, and real-time imaging properties. ,,− In these methods, o-diaminophenyl group, a fluorescence quencher and a NO trapper, was linked to the probe, which can completely quench the fluorescence of probe by PET process . After reacting with NO to form corresponding benzotriazole derivative, the fluorescence turns on dramatically as the PET process is inhibited.…”