Molecular interface of S100A8 with cytochrome b558 and NADPH oxidase activation

Berthier, Sylvie; Nguyen, Minh Vu Chong; Baillet, Athan; Hograindleur, Marc-André; Paclet, Marie- Hélène; Polack, Benoît; Morel, Françoise

doi:10.1371/journal.pone.0040277

Reference : Molecular interface of S100A8 with cytochrome b558 and NADPH oxidase activation


	Document type :	Scientific journals : Article
	Discipline(s) :	Life sciences : Biochemistry, biophysics & molecular biology
	To cite this reference:	http://hdl.handle.net/10993/27258

Title :	Molecular interface of S100A8 with cytochrome b558 and NADPH oxidase activation
Language :	English
Author, co-author :	Berthier, Sylvie [Groupe de Recherche et d'Etude du Processus Inflammatoire, Laboratoire Aging Imaging Modeling(AGIM), Formation de Recherche en evolution (FRE) Centre National de la Recherche Scientifique CNRS 3405, Université Joseph Fourier UJF, Grenoble, France]
	Nguyen, Minh Vu Chong [University of Luxembourg > Faculty of Science, Technology and Communication (FSTC) > Life Science Research Unit]
	Baillet, Athan [Groupe de Recherche et d'Etude du Processus Inflammatoire, Laboratoire Aging Imaging Modeling(AGIM), Formation de Recherche en evolution (FRE) Centre National de la Recherche Scientifique CNRS 3405, Université Joseph Fourier UJF, Grenoble, France, Clinic of Rheumatology, Centre Hospitalier Universitaire (CHU), Grenoble, France]
	Hograindleur, Marc-André [Groupe de Recherche et d'Etude du Processus Inflammatoire, Laboratoire Aging Imaging Modeling(AGIM), Formation de Recherche en evolution (FRE) Centre National de la Recherche Scientifique CNRS 3405, Université Joseph Fourier UJF, Grenoble, France]
	Paclet, Marie- Hélène [Groupe de Recherche et d'Etude du Processus Inflammatoire, Laboratoire Aging Imaging Modeling(AGIM), Formation de Recherche en evolution (FRE) Centre National de la Recherche Scientifique CNRS 3405, Université Joseph Fourier UJF, Grenoble, France, Laboratoire des Enzymes et des Protéines, Centre Hospitalier Universitaire (CHU), Grenoble, France, Institut de Biologie et Pathologie, Centre Hospitalier Universitaire (CHU), Grenoble, France]
	Polack, Benoît [Institut de Biologie et Pathologie, Centre Hospitalier Universitaire (CHU), Grenoble, France, Techniques de l'Ingénierie Médicale et de la Complexité-Informatique, Mathématiques et Applications de Grenoble (TIMC-IMAG) Unité Mixte de Recherche (UMR), 5525 Centre National de la Recherche Scientifique (CNRS), Université Joseph Fourier UJF, Grenoble, France]
	Morel, Françoise [Groupe de Recherche et d'Etude du Processus Inflammatoire, Laboratoire Aging Imaging Modeling(AGIM), Formation de Recherche en evolution (FRE) Centre National de la Recherche Scientifique CNRS 3405, Université Joseph Fourier UJF, Grenoble, France]
Publication date :	2012
Journal title :	PLoS ONE
Volume :	7
Issue/season :	7
Peer reviewed :	Yes (verified by ORBi^lu)
Audience :	International
ISSN :	19326203
Keywords :	[en] ExoS toxin ; Escherichia coli ; Pseudomonas aeruginosa ; Amino Acid Sequence ; Antibodies, Monoclonal ; Bacterial Secretion Systems ; Calgranulin A ; Calgranulin B ; Cell-Free System ; Cross-Linking Reagents ; Cytochrome b Group ; Cytosol ; Enzyme Activation ; Herpesvirus 4, Human ; Humans ; Lymphocytes ; Molecular Sequence Data ; NADPH Oxidase ; Neutrophils ; Protein Binding ; Protein Structure, Tertiary ; Protein Transport ; Pseudomonas aeruginosa ; Recombinant Proteins ; Structure-Activity Relationship
Abstract :	[en] S100A8 and S100A9 are two calcium binding Myeloid Related Proteins, and important mediators of inflammatory diseases. They were recently introduced as partners for phagocyte NADPH oxidase regulation. However, the precise mechanism of their interaction remains elusive. We had for aim (i) to evaluate the impact of S100 proteins on NADPH oxidase activity; (ii) to characterize molecular interaction of either S100A8, S100A9, or S100A8/S100A9 heterocomplex with cytochrome b558; and (iii) to determine the S100A8 consensus site involved in cytochrome b558/S100 interface. Recombinant full length or S100A9-A8 truncated chimera proteins and ExoS-S100 fusion proteins were expressed in E. coli and in P. aeruginosa respectively. Our results showed that S100A8 is the functional partner for NADPH oxidase activation contrary to S100A9, however, the loading with calcium and a combination with phosphorylated S100A9 are essential in vivo. Endogenous S100A9 and S100A8 colocalize in differentiated and PMA stimulated PLB985 cells, with Nox2/gp91phox and p22phox. Recombinant S100A8, loaded with calcium and fused with the first 129 or 54 N-terminal amino acid residues of the P. aeruginosa ExoS toxin, induced a similar oxidase activation in vitro, to the one observed with S100A8 in the presence of S100A9 in vivo. This suggests that S100A8 is the essential component of the S100A9/S100A8 heterocomplex for oxidase activation. In this context, recombinant full-length rS100A9-A8 and rS100A9-A8 truncated 90 chimera proteins as opposed to rS100A9-A8 truncated 86 and rS100A9-A8 truncated 57 chimeras, activate the NADPH oxidase function of purified cytochrome b558 suggesting that the C-terminal region of S100A8 is directly involved in the molecular interface with the hemoprotein. The data point to four strategic 87HEES90 amino acid residues of the S100A8 C-terminal sequence that are involved directly in the molecular interaction with cytochrome b558 and then in the phagocyte NADPH oxidase activation. © 2012 Berthier et al.
Permalink :	http://hdl.handle.net/10993/27258
DOI :	10.1371/journal.pone.0040277

File(s) associated to this reference

Fulltext file(s):

	File	Commentary	Version	Size	Access
Open access	Molecular Interface of S100A8.PDF		Publisher postprint	2.43 MB	View/Open

All documents in ORBi^lu are protected by a user license.

University of Luxembourg Library | Feedback | Legal notices