Solution structure analysis of the periplasmic region of bacterial flagellar motor stators by small angle X-ray scattering

Liew, CW; Hynson, RM; Ganuelas, LA; Shah-Mohammadi, N; Duff, AP; Kojima, S; Homma, M; Lee, LK

Solution structure analysis of the periplasmic region of bacterial flagellar motor stators by small angle X-ray scattering

Liew, CW Hynson, RM Ganuelas, LA Shah-Mohammadi, N Duff, AP Kojima, S Homma, M Lee, LK

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Publication Type:: Journal Article
Citation:: Biochemical and Biophysical Research Communications, 2018, 495 (2), pp. 1614 - 1619
Issue Date:: 2018-01-08

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Field	Value	Language
dc.contributor.author	Liew, CW	en_US
dc.contributor.author	Hynson, RM	en_US
dc.contributor.author	Ganuelas, LA	en_US
dc.contributor.author	Shah-Mohammadi, N	en_US
dc.contributor.author	Duff, AP	en_US
dc.contributor.author	Kojima, S	en_US
dc.contributor.author	Homma, M	en_US
dc.contributor.author	Lee, LK	en_US
dc.date.available	2017-11-28	en_US
dc.date.issued	2018-01-08	en_US
dc.identifier.citation	Biochemical and Biophysical Research Communications, 2018, 495 (2), pp. 1614 - 1619	en_US
dc.identifier.issn	0006-291X	en_US
dc.identifier.uri	http://hdl.handle.net/10453/131008
dc.description.abstract	© 2017 Elsevier Inc. The bacterial flagellar motor drives the rotation of helical flagellar filaments to propel bacteria through viscous media. It consists of a dynamic population of mechanosensitive stators that are embedded in the inner membrane and activate in response to external load. This entails assembly around the rotor, anchoring to the peptidoglycan layer to counteract torque from the rotor and opening of a cation channel to facilitate an influx of cations, which is converted into mechanical rotation. Stator complexes are comprised of four copies of an integral membrane A subunit and two copies of a B subunit. Each B subunit includes a C-terminal OmpA-like peptidoglycan-binding (PGB) domain. This is thought to be linked to a single N-terminal transmembrane helix by a long unstructured peptide, which allows the PGB domain to bind to the peptidoglycan layer during stator anchoring. The high-resolution crystal structures of flagellar motor PGB domains from Salmonella enterica (MotBC2) and Vibrio alginolyticus (PomBC5) have previously been elucidated. Here, we use small-angle X-ray scattering (SAXS). We show that unlike MotBC2, the dimeric conformation of the PomBC5 in solution differs to its crystal structure, and explore the functional relevance by characterising gain-of-function mutants as well as wild-type constructs of various lengths. These provide new insight into the conformational diversity of flagellar motor PGB domains and experimental verification of their overall topology.	en_US
dc.relation.ispartof	Biochemical and Biophysical Research Communications	en_US
dc.relation.isbasedon	10.1016/j.bbrc.2017.11.194	en_US
dc.subject.classification	Biochemistry & Molecular Biology	en_US
dc.subject.mesh	Flagella	en_US
dc.subject.mesh	Salmonella enterica	en_US
dc.subject.mesh	Vibrio alginolyticus	en_US
dc.subject.mesh	Bacterial Proteins	en_US
dc.subject.mesh	Bacterial Outer Membrane Proteins	en_US
dc.subject.mesh	Recombinant Proteins	en_US
dc.subject.mesh	Solutions	en_US
dc.subject.mesh	X-Ray Diffraction	en_US
dc.subject.mesh	Amino Acid Sequence	en_US
dc.subject.mesh	Protein Structure, Quaternary	en_US
dc.subject.mesh	Sequence Homology, Amino Acid	en_US
dc.subject.mesh	Models, Molecular	en_US
dc.subject.mesh	Molecular Motor Proteins	en_US
dc.subject.mesh	Scattering, Small Angle	en_US
dc.subject.mesh	Protein Domains	en_US
dc.title	Solution structure analysis of the periplasmic region of bacterial flagellar motor stators by small angle X-ray scattering	en_US
dc.type	Journal Article
utslib.citation.volume	2	en_US
utslib.citation.volume	495	en_US
utslib.for	0304 Medicinal and Biomolecular Chemistry	en_US
utslib.for	0601 Biochemistry and Cell Biology	en_US
utslib.for	1101 Medical Biochemistry and Metabolomics	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
utslib.copyright.status	closed_access
pubs.issue	2	en_US
pubs.publication-status	Published	en_US
pubs.volume	495	en_US

Abstract:

© 2017 Elsevier Inc. The bacterial flagellar motor drives the rotation of helical flagellar filaments to propel bacteria through viscous media. It consists of a dynamic population of mechanosensitive stators that are embedded in the inner membrane and activate in response to external load. This entails assembly around the rotor, anchoring to the peptidoglycan layer to counteract torque from the rotor and opening of a cation channel to facilitate an influx of cations, which is converted into mechanical rotation. Stator complexes are comprised of four copies of an integral membrane A subunit and two copies of a B subunit. Each B subunit includes a C-terminal OmpA-like peptidoglycan-binding (PGB) domain. This is thought to be linked to a single N-terminal transmembrane helix by a long unstructured peptide, which allows the PGB domain to bind to the peptidoglycan layer during stator anchoring. The high-resolution crystal structures of flagellar motor PGB domains from Salmonella enterica (MotBC2) and Vibrio alginolyticus (PomBC5) have previously been elucidated. Here, we use small-angle X-ray scattering (SAXS). We show that unlike MotBC2, the dimeric conformation of the PomBC5 in solution differs to its crystal structure, and explore the functional relevance by characterising gain-of-function mutants as well as wild-type constructs of various lengths. These provide new insight into the conformational diversity of flagellar motor PGB domains and experimental verification of their overall topology.

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http://hdl.handle.net/10453/131008