Structural and biochemical characterization of Acinetobacter baumannii ZnuA.

Alquethamy, S; Ganio, K; Luo, Z; Hossain, SI; Hayes, AJ; Ve, T; Davies, MR; Deplazes, E; Kobe, B; McDevitt, CA

Structural and biochemical characterization of Acinetobacter baumannii ZnuA.

Alquethamy, S Ganio, K Luo, Z Hossain, SI Hayes, AJ Ve, T Davies, MR Deplazes, E

Kobe, B McDevitt, CA

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Publisher:: Elsevier
Publication Type:: Journal Article
Citation:: Journal of Inorganic Biochemistry, 2022, 231, pp. 1-11
Issue Date:: 2022-06

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Field	Value	Language
dc.contributor.author	Alquethamy, S
dc.contributor.author	Ganio, K
dc.contributor.author	Luo, Z
dc.contributor.author	Hossain, SI
dc.contributor.author	Hayes, AJ
dc.contributor.author	Ve, T
dc.contributor.author	Davies, MR
dc.contributor.author	Deplazes, E https://orcid.org/0000-0003-2052-5536
dc.contributor.author	Kobe, B
dc.contributor.author	McDevitt, CA
dc.date.accessioned	2022-11-09T04:41:03Z
dc.date.available	2022-03-06
dc.date.available	2022-11-09T04:41:03Z
dc.date.issued	2022-06
dc.identifier.citation	Journal of Inorganic Biochemistry, 2022, 231, pp. 1-11
dc.identifier.issn	0162-0134
dc.identifier.issn	1873-3344
dc.identifier.uri	http://hdl.handle.net/10453/163372
dc.description.abstract	Acinetobacter baumannii is a Gram-negative nosocomial pathogen associated with significant disease. Crucial to the survival and pathogenesis of A. baumannii is the ability to acquire essential micronutrients such as Zn(II). Recruitment of Zn(II) by A. baumannii is mediated, at least in part, by the periplasmic solute-binding protein ZnuA and the ATP-binding cassette transporter ZnuBC. Here, we combined genomic, biochemical, and structural approaches to characterize A. baumannii AB5075_UW ZnuA. Bioinformatic analyses using a diverse collection of A. baumannii genomes determined that ZnuA is highly conserved, with the binding site comprised by three strictly conserved histidine residues. The structure of metal-free ZnuA was determined at 2.1 Å resolution, with molecular dynamics analyses revealing loop α2β2, which harbors the putative Zn(II)-coordinating residue His41, to be highly mobile in the metal-free state. The contribution of the putative binding site histidine residues to Zn(II) interaction was further probed by mutagenesis. Analysis of ZnuA mutant variants was performed by quantitative metal binding assays, differential scanning fluorimetry, and affinity measurements, which showed that all three histidine residues contributed to Zn(II)-recruitment, albeit to different extents. Collectively, these analyses provide insight into the mechanism of Zn(II)-binding by A. baumannii ZnuA and expand our understanding of the functional diversity of Zn(II)-recruiting proteins.
dc.format	Print-Electronic
dc.language	eng
dc.publisher	Elsevier
dc.relation	http://purl.org/au-research/grants/nhmrc/1122582
dc.relation.ispartof	Journal of Inorganic Biochemistry
dc.relation.isbasedon	10.1016/j.jinorgbio.2022.111787
dc.rights	info:eu-repo/semantics/closedAccess
dc.subject	0302 Inorganic Chemistry, 0307 Theoretical and Computational Chemistry, 0399 Other Chemical Sciences
dc.subject.classification	Inorganic & Nuclear Chemistry
dc.subject.mesh	Acinetobacter baumannii
dc.subject.mesh	ATP-Binding Cassette Transporters
dc.subject.mesh	Bacterial Proteins
dc.subject.mesh	Histidine
dc.subject.mesh	Models, Molecular
dc.subject.mesh	Zinc
dc.subject.mesh	ATP-Binding Cassette Transporters
dc.subject.mesh	Acinetobacter baumannii
dc.subject.mesh	Bacterial Proteins
dc.subject.mesh	Histidine
dc.subject.mesh	Models, Molecular
dc.subject.mesh	Zinc
dc.subject.mesh	Acinetobacter baumannii
dc.subject.mesh	Zinc
dc.subject.mesh	Histidine
dc.subject.mesh	Bacterial Proteins
dc.subject.mesh	ATP-Binding Cassette Transporters
dc.subject.mesh	Models, Molecular
dc.title	Structural and biochemical characterization of Acinetobacter baumannii ZnuA.
dc.type	Journal Article
utslib.citation.volume	231
utslib.location.activity	United States
utslib.for	0302 Inorganic Chemistry
utslib.for	0307 Theoretical and Computational Chemistry
utslib.for	0399 Other Chemical Sciences
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology
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/Faculty of Engineering and Information Technology/School of Mechanical and Mechatronic Engineering
utslib.copyright.status	closed_access	*
pubs.consider-herdc	false
dc.date.updated	2022-11-09T04:40:55Z
pubs.publication-status	Published
pubs.volume	231

Abstract:

Acinetobacter baumannii is a Gram-negative nosocomial pathogen associated with significant disease. Crucial to the survival and pathogenesis of A. baumannii is the ability to acquire essential micronutrients such as Zn(II). Recruitment of Zn(II) by A. baumannii is mediated, at least in part, by the periplasmic solute-binding protein ZnuA and the ATP-binding cassette transporter ZnuBC. Here, we combined genomic, biochemical, and structural approaches to characterize A. baumannii AB5075_UW ZnuA. Bioinformatic analyses using a diverse collection of A. baumannii genomes determined that ZnuA is highly conserved, with the binding site comprised by three strictly conserved histidine residues. The structure of metal-free ZnuA was determined at 2.1 Å resolution, with molecular dynamics analyses revealing loop α2β2, which harbors the putative Zn(II)-coordinating residue His41, to be highly mobile in the metal-free state. The contribution of the putative binding site histidine residues to Zn(II) interaction was further probed by mutagenesis. Analysis of ZnuA mutant variants was performed by quantitative metal binding assays, differential scanning fluorimetry, and affinity measurements, which showed that all three histidine residues contributed to Zn(II)-recruitment, albeit to different extents. Collectively, these analyses provide insight into the mechanism of Zn(II)-binding by A. baumannii ZnuA and expand our understanding of the functional diversity of Zn(II)-recruiting proteins.

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http://hdl.handle.net/10453/163372