A DNA-based fluorescent probe maps NOS3 activity with subcellular spatial resolution.

Nature Chemical Biology
Maulik S JaniYamuna Krishnan

Abstract

Nitric oxide synthase 3 (NOS3) produces the gasotransmitter nitric oxide (NO), which drives critical cellular signaling pathways by S-nitrosylating target proteins. Endogenous NOS3 resides at two distinct subcellular locations: the plasma membrane and the trans-Golgi network (TGN). However, NO generation arising from the activities of both these pools of NOS3 and its relative contribution to physiology or disease is not yet resolvable. We describe a fluorescent DNA-based probe technology, NOckout, that can be targeted either to the plasma membrane or the TGN, where it can quantitatively map the activities of endogenous NOS3 at these locations in live cells. We found that, although NOS3 at the Golgi is tenfold less active than at the plasma membrane, its activity is essential for the structural integrity of the Golgi. The newfound ability to spatially map NOS3 activity provides a platform to discover selective regulators of the distinct pools of NOS3.

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Citations

May 24, 2020·Nature Chemical Biology·Emrah ErogluRoland Malli
Jun 20, 2020·Proceedings of the National Academy of Sciences of the United States of America·Aneesh T VeetilYamuna Krishnan
Oct 1, 2020·Angewandte Chemie·Obianuju A NwokoloIrina V Nesterova
Nov 4, 2020·Nature Nanotechnology·Anand SaminathanYamuna Krishnan
Mar 17, 2021·Trends in Biotechnology·Andreas SchoenitKerstin Göpfrich
Dec 5, 2020·ACS Central Science·Yamuna KrishnanMaulik S Jani
May 25, 2021·Chemical Society Reviews·Hao-Ran JiaFu-Gen Wu
Jul 29, 2021·ELife·Kasturi ChakrabortyYamuna Krishnan
Jun 24, 2020·Journal of the American Chemical Society·Sasha B EbrahimiChad A Mirkin
Oct 6, 2021·Proceedings of the National Academy of Sciences of the United States of America·Bhavyashree SureshYamuna Krishnan
Oct 28, 2021·Chemical Communications : Chem Comm·Sumit Kumar Pramanik, Amitva Das

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Datasets Mentioned

BETA
GM130
GM-130

Methods Mentioned

BETA
electrophoresis
Fluorescence
fluorescence imaging

Software Mentioned

NOckout
Fiji

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