Description
Nitric oxide (NO) exerts multiple biological functions through autocrine and paracrine signaling pathways, particularly in the regulation of vascular tone, neurotransmission, acute and chronic inflammation, and host defense against pathogens, including viruses, bacteria, and fungi. Following real-time NO fluctuations in concentration at the single-cell level is a challenging task due to its low concentration, short lifetime, and high reactivity with a variety of reactive oxygen species.
Only a few genetically encoded fluorescent sensors (GES) have been so far developed and proposed for monitoring intracellular NO levels, but their complexity, large size and multi-domain structure raise concerns about their reliability. These limits highlight the need for the development of sensitive yet simpler GES for NO monitoring.
This project exploits our seminal observation that the blue-emitting fluorescent protein mTagBFP2, undergoes a reduction in fluorescence quantum yield and lifetime upon exposure to μM NO concentrations, an effect that has been correlated with S-nitrosylation of Cys residues.
Taking advantage of a multidisciplinary team with internationally-recognized expertise in chemical physics of biological systems, protein design, and engineering, time-resolved spectroscopy, and fluorescence microscopy, this project proposes to explore fluorescent proteins (FP) as simple NO sensors to obtain a molecular platform of GES suitable for fluorescence imaging, including fluorescence lifetime imaging (FLIM).
Project References
PNRR-M4C2- I1.1- Avviso MUR n. 1409 del 14-09-2022 – Bando PRIN 2022 PNRR - Settore ERC PE4 Titolo Progetto: A molecular platform for intracellular nitric oxide sensing - Codice Progetto P2022F4WR8 - Codice CUP D53D23016840001 - Finanziato dall’Unione Europea – NextGenerationEU
Team
