Functional Characterization of the N-Acetylmuramyl-l-Alanine Amidase, Ami1, from Mycobacterium abscessus.

Küssau, T; Van Wyk, N; Johansen, MD; Alsarraf, HMAB; Neyret, A; Hamela, C; Sørensen, KK; Thygesen, MB; Beauvineau, C; Kremer, L; Blaise, M

Functional Characterization of the N-Acetylmuramyl-l-Alanine Amidase, Ami1, from Mycobacterium abscessus.

Küssau, T Van Wyk, N Johansen, MD Alsarraf, HMAB Neyret, A Hamela, C Sørensen, KK Thygesen, MB Beauvineau, C Kremer, L Blaise, M

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Publisher:: MDPI AG
Publication Type:: Journal Article
Citation:: Cells, 2020, 9, (11)
Issue Date:: 2020-11-04

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Field	Value	Language
dc.contributor.author	Küssau, T
dc.contributor.author	Van Wyk, N
dc.contributor.author	Johansen, MD
dc.contributor.author	Alsarraf, HMAB
dc.contributor.author	Neyret, A
dc.contributor.author	Hamela, C
dc.contributor.author	Sørensen, KK
dc.contributor.author	Thygesen, MB
dc.contributor.author	Beauvineau, C
dc.contributor.author	Kremer, L
dc.contributor.author	Blaise, M
dc.date.accessioned	2020-11-27T02:38:29Z
dc.date.available	2020-10-29
dc.date.available	2020-11-27T02:38:29Z
dc.date.issued	2020-11-04
dc.identifier.citation	Cells, 2020, 9, (11)
dc.identifier.issn	2073-4409
dc.identifier.issn	2073-4409
dc.identifier.uri	http://hdl.handle.net/10453/144402
dc.description.abstract	Peptidoglycan (PG) is made of a polymer of disaccharides organized as a three-dimensional mesh-like network connected together by peptidic cross-links. PG is a dynamic structure that is essential for resistance to environmental stressors. Remodeling of PG occurs throughout the bacterial life cycle, particularly during bacterial division and separation into daughter cells. Numerous autolysins with various substrate specificities participate in PG remodeling. Expression of these enzymes must be tightly regulated, as an excess of hydrolytic activity can be detrimental for the bacteria. In non-tuberculous mycobacteria such as Mycobacterium abscessus, the function of PG-modifying enzymes has been poorly investigated. In this study, we characterized the function of the PG amidase, Ami1 from M. abscessus. An ami1 deletion mutant was generated and the phenotypes of the mutant were evaluated with respect to susceptibility to antibiotics and virulence in human macrophages and zebrafish. The capacity of purified Ami1 to hydrolyze muramyl-dipeptide was demonstrated in vitro. In addition, the screening of a 9200 compounds library led to the selection of three compounds inhibiting Ami1 in vitro. We also report the structural characterization of Ami1 which, combined with in silico docking studies, allows us to propose a mode of action for these inhibitors.
dc.format	Electronic
dc.language	eng
dc.publisher	MDPI AG
dc.relation.ispartof	Cells
dc.relation.isbasedon	10.3390/cells9112410
dc.rights	info:eu-repo/semantics/openAccess
dc.title	Functional Characterization of the N-Acetylmuramyl-l-Alanine Amidase, Ami1, from Mycobacterium abscessus.
dc.type	Journal Article
utslib.citation.volume	9
utslib.location.activity	Switzerland
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
pubs.organisational-group	/University of Technology Sydney
utslib.copyright.status	open_access	*
pubs.consider-herdc	false
dc.date.updated	2020-11-27T02:38:14Z
pubs.issue	11
pubs.publication-status	Published
pubs.volume	9
utslib.citation.issue	11

Abstract:

Peptidoglycan (PG) is made of a polymer of disaccharides organized as a three-dimensional mesh-like network connected together by peptidic cross-links. PG is a dynamic structure that is essential for resistance to environmental stressors. Remodeling of PG occurs throughout the bacterial life cycle, particularly during bacterial division and separation into daughter cells. Numerous autolysins with various substrate specificities participate in PG remodeling. Expression of these enzymes must be tightly regulated, as an excess of hydrolytic activity can be detrimental for the bacteria. In non-tuberculous mycobacteria such as Mycobacterium abscessus, the function of PG-modifying enzymes has been poorly investigated. In this study, we characterized the function of the PG amidase, Ami1 from M. abscessus. An ami1 deletion mutant was generated and the phenotypes of the mutant were evaluated with respect to susceptibility to antibiotics and virulence in human macrophages and zebrafish. The capacity of purified Ami1 to hydrolyze muramyl-dipeptide was demonstrated in vitro. In addition, the screening of a 9200 compounds library led to the selection of three compounds inhibiting Ami1 in vitro. We also report the structural characterization of Ami1 which, combined with in silico docking studies, allows us to propose a mode of action for these inhibitors.

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http://hdl.handle.net/10453/144402