The NOX Family of Proteins Is Also Present in Bacteria

ABSTRACT Transmembrane NADPH oxidase (NOX) enzymes have been so far only characterized in eukaryotes. In most of these organisms, they reduce molecular oxygen to superoxide and, depending on the presence of additional domains, are called NOX or dual oxidases (DUOX). Reactive oxygen species (ROS), in...

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Autores principales:	Christine Hajjar, Mickaël V. Cherrier, Gaëtan Dias Mirandela, Isabelle Petit-Hartlein, Marie José Stasia, Juan C. Fontecilla-Camps, Franck Fieschi, Jérôme Dupuy
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Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	Streptococcus pneumoniae biochemistry electron transport flavoenzymes membrane proteins metalloenzymes Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/6cb90c7e1136436e82aedc8d6d255758
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spelling	oai:doaj.org-article:6cb90c7e1136436e82aedc8d6d2557582021-11-15T15:51:55ZThe NOX Family of Proteins Is Also Present in Bacteria10.1128/mBio.01487-172150-7511https://doaj.org/article/6cb90c7e1136436e82aedc8d6d2557582017-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01487-17https://doaj.org/toc/2150-7511ABSTRACT Transmembrane NADPH oxidase (NOX) enzymes have been so far only characterized in eukaryotes. In most of these organisms, they reduce molecular oxygen to superoxide and, depending on the presence of additional domains, are called NOX or dual oxidases (DUOX). Reactive oxygen species (ROS), including superoxide, have been traditionally considered accidental toxic by-products of aerobic metabolism. However, during the last decade it has become evident that both O2•− and H2O2 are key players in complex signaling networks and defense. A well-studied example is the production of O2•− during the bactericidal respiratory burst of phagocytes; this production is catalyzed by NOX2. Here, we devised and applied a novel algorithm to search for additional NOX genes in genomic databases. This procedure allowed us to discover approximately 23% new sequences from bacteria (in relation to the number of NOX-related sequences identified by the authors) that we have added to the existing eukaryotic NOX family and have used to build an expanded phylogenetic tree. We cloned and overexpressed the identified nox gene from Streptococcus pneumoniae and confirmed that it codes for an NADPH oxidase. The membrane of the S. pneumoniae NOX protein (SpNOX) shares many properties with its eukaryotic counterparts, such as affinity for NADPH and flavin adenine dinucleotide, superoxide dismutase and diphenylene iodonium inhibition, cyanide resistance, oxygen consumption, and superoxide production. Traditionally, NOX enzymes in eukaryotes are related to functions linked to multicellularity. Thus, the discovery of a large family of NOX-related enzymes in the bacterial world brings up fascinating questions regarding their role in this new biological context. IMPORTANCE NADPH oxidase (NOX) enzymes have not yet been reported in bacteria. Here, we carried out computational and experimental studies to provide the first characterization of a prokaryotic NOX. Out of 996 prokaryotic proteins showing NOX signatures, we initially selected, cloned, and overexpressed four of them. Subsequently, and based on preliminary testing, we concentrated our efforts on Streptococcus SpNOX, which shares many biochemical characteristics with NOX2, the referent model of NOX enzymes. Our work makes possible, for the first time, the study of pure forms of this important family of enzymes, allowing for biophysical and molecular characterization in an unprecedented way. Similar advances regarding other membrane protein families have led to new structures, further mechanistic studies, and the improvement of inhibitors. In addition, biological functions of these newly described bacterial enzymes will be certainly discovered in the near future.Christine HajjarMickaël V. CherrierGaëtan Dias MirandelaIsabelle Petit-HartleinMarie José StasiaJuan C. Fontecilla-CampsFranck FieschiJérôme DupuyAmerican Society for MicrobiologyarticleStreptococcus pneumoniaebiochemistryelectron transportflavoenzymesmembrane proteinsmetalloenzymesMicrobiologyQR1-502ENmBio, Vol 8, Iss 6 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Streptococcus pneumoniae biochemistry electron transport flavoenzymes membrane proteins metalloenzymes Microbiology QR1-502
spellingShingle	Streptococcus pneumoniae biochemistry electron transport flavoenzymes membrane proteins metalloenzymes Microbiology QR1-502 Christine Hajjar Mickaël V. Cherrier Gaëtan Dias Mirandela Isabelle Petit-Hartlein Marie José Stasia Juan C. Fontecilla-Camps Franck Fieschi Jérôme Dupuy The NOX Family of Proteins Is Also Present in Bacteria
description	ABSTRACT Transmembrane NADPH oxidase (NOX) enzymes have been so far only characterized in eukaryotes. In most of these organisms, they reduce molecular oxygen to superoxide and, depending on the presence of additional domains, are called NOX or dual oxidases (DUOX). Reactive oxygen species (ROS), including superoxide, have been traditionally considered accidental toxic by-products of aerobic metabolism. However, during the last decade it has become evident that both O2•− and H2O2 are key players in complex signaling networks and defense. A well-studied example is the production of O2•− during the bactericidal respiratory burst of phagocytes; this production is catalyzed by NOX2. Here, we devised and applied a novel algorithm to search for additional NOX genes in genomic databases. This procedure allowed us to discover approximately 23% new sequences from bacteria (in relation to the number of NOX-related sequences identified by the authors) that we have added to the existing eukaryotic NOX family and have used to build an expanded phylogenetic tree. We cloned and overexpressed the identified nox gene from Streptococcus pneumoniae and confirmed that it codes for an NADPH oxidase. The membrane of the S. pneumoniae NOX protein (SpNOX) shares many properties with its eukaryotic counterparts, such as affinity for NADPH and flavin adenine dinucleotide, superoxide dismutase and diphenylene iodonium inhibition, cyanide resistance, oxygen consumption, and superoxide production. Traditionally, NOX enzymes in eukaryotes are related to functions linked to multicellularity. Thus, the discovery of a large family of NOX-related enzymes in the bacterial world brings up fascinating questions regarding their role in this new biological context. IMPORTANCE NADPH oxidase (NOX) enzymes have not yet been reported in bacteria. Here, we carried out computational and experimental studies to provide the first characterization of a prokaryotic NOX. Out of 996 prokaryotic proteins showing NOX signatures, we initially selected, cloned, and overexpressed four of them. Subsequently, and based on preliminary testing, we concentrated our efforts on Streptococcus SpNOX, which shares many biochemical characteristics with NOX2, the referent model of NOX enzymes. Our work makes possible, for the first time, the study of pure forms of this important family of enzymes, allowing for biophysical and molecular characterization in an unprecedented way. Similar advances regarding other membrane protein families have led to new structures, further mechanistic studies, and the improvement of inhibitors. In addition, biological functions of these newly described bacterial enzymes will be certainly discovered in the near future.
format	article
author	Christine Hajjar Mickaël V. Cherrier Gaëtan Dias Mirandela Isabelle Petit-Hartlein Marie José Stasia Juan C. Fontecilla-Camps Franck Fieschi Jérôme Dupuy
author_facet	Christine Hajjar Mickaël V. Cherrier Gaëtan Dias Mirandela Isabelle Petit-Hartlein Marie José Stasia Juan C. Fontecilla-Camps Franck Fieschi Jérôme Dupuy
author_sort	Christine Hajjar
title	The NOX Family of Proteins Is Also Present in Bacteria
title_short	The NOX Family of Proteins Is Also Present in Bacteria
title_full	The NOX Family of Proteins Is Also Present in Bacteria
title_fullStr	The NOX Family of Proteins Is Also Present in Bacteria
title_full_unstemmed	The NOX Family of Proteins Is Also Present in Bacteria
title_sort	nox family of proteins is also present in bacteria
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/6cb90c7e1136436e82aedc8d6d255758
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The NOX Family of Proteins Is Also Present in Bacteria

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