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FEBS J. 2019 Nov 30. doi: 10.1111/febs.15160. [Epub ahead of print]

On the mechanism of calcium-dependent activation of NADPH oxidase 5 (NOX5).

Author information

1
Department of Biology and Biotechnology "Lazzaro Spallanzani", University of Pavia, Italy.
2
UK Dementia Research Institute at King's College London, UK.
3
The Wohl Institute at King's College London, UK.
4
The Crick Institute, London, UK.
5
Institute of Biotechnology, Czech Academy of Sciences, Vestec, Czech Republic.
6
Institute of Microbiology, Czech Academy of Sciences, Prague, Czech Republic.

Abstract

It is now accepted that reactive oxygen species (ROS) are not only dangerous oxidative agents but also chemical mediators of the redox cell signaling and innate immune response. A central role in ROS-controlled production is played by the NADPH oxidases (NOXs), a group of seven membrane-bound enzymes (NOX1-5 and DUOX1-2) whose unique function is to produce ROS. Here, we describe the regulation of NOX5, a widespread family member present in cyanobacteria, protists, plants, fungi, and the animal kingdom. We show that the calmodulin-like regulatory EF-domain of NOX5 is partially unfolded and detached from the rest of the protein in the absence of calcium. In the presence of calcium, the C-terminal lobe of the EF-domain acquires an ordered and more compact structure that enables its binding to the enzyme dehydrogenase (DH) domain. Our spectroscopic and mutagenesis studies further identified a set of conserved aspartate residues in the DH domain that are essential for NOX5 activation. Altogether, our work shows that calcium induces an unfolded-to-folded transition of the EF-domain that promotes direct interaction with a conserved regulatory region, resulting in NOX5 activation.

KEYWORDS:

EF-hands; NMR; calcium activation; enzyme; structure

PMID:
31785178
DOI:
10.1111/febs.15160

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