An integrated pharmacological, structural, and genetic analysis of extracellular versus intracellular ROS production in neutrophils

Item Type:	Article
Title:	An integrated pharmacological, structural, and genetic analysis of extracellular versus intracellular ROS production in neutrophils
Creators Name:	Ellson, C.D. and Goretti Riça, I. and Kim, J.S. and Huang, Y.M.M. and Lim, D. and Mitra, T. and Hsu, A. and Wei, E.X. and Barrett, C.D. and Otterbein, L.E. and Hauser, C.J. and Wahl, M. and Delbrück, H. and Heinemann, U. and Oschkinat, H. and Chang, C.E.A. and Yaffe, M.B.
Abstract:	The neutrophil NADPH oxidase produces both intracellular and extracellular reactive oxygen species (ROS). Although oxidase activity is essential for microbial killing, and ROS can act as signaling molecules in the inflammatory process, excessive extracellular ROS directly contributes to inflammatory tissue damage, as well as to cancer progression and immune dysregulation in the tumor microenvironment. How specific signaling pathways contribute to ROS localization is unclear. Here we used a systems pharmacology approach to identify the specific Class I PI3-K isoform p110β, and PLD1, but not PLD2, as critical regulators of extracellular, but not intracellular ROS production in primary neutrophils. Combined crystallographic and molecular dynamics analysis of the PX domain of the oxidase component p47phox, which binds the lipid products of PI 3-K and PLD, was used to clarify the membrane-binding mechanism and guide the design of mutant mice whose p47phox is unable to bind 3-phosphorylated inositol phospholipids. Neutrophils from these K43A mutant animals were specifically deficient in extracellular, but not intracellular, ROS production, and showed increased dependency on signaling through the remaining PLD1 arm. These findings identify the PX domain of p47phox as a critical integrator of PLD1 and p110β signaling for extracellular ROS production, and as a potential therapeutic target for modulating tissue damage and extracellular signaling during inflammation.
Keywords:	Inflammation, NADPH Oxidases, Neutrophils, Reactive Oxygen Species, Signal Transduction, Animals, Mice
Source:	Journal of Molecular Biology
ISSN:	0022-2836
Publisher:	Elsevier
Volume:	434
Number:	9
Page Range:	167533
Date:	15 May 2022
Official Publication:	https://doi.org/10.1016/j.jmb.2022.167533
PubMed:	View item in PubMed

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