Structures of the Ultra-High-Affinity Protein-Protein Complexes of Pyocins S2 and AP41 and Their Cognate Immunity Proteins from Pseudomonas aeruginosa

Joshi, A; Grinter, R; Josts, I; Chen, S; Wojdyla, JA; Lowe, ED; Kaminska, R; Sharp, C; McCaughey, L; Roszak, AW; Cogdell, RJ; Byron, O; Walker, D; Kleanthous, C

Structures of the Ultra-High-Affinity Protein-Protein Complexes of Pyocins S2 and AP41 and Their Cognate Immunity Proteins from Pseudomonas aeruginosa

Joshi, A Grinter, R Josts, I Chen, S Wojdyla, JA Lowe, ED Kaminska, R Sharp, C McCaughey, L

Roszak, AW Cogdell, RJ Byron, O Walker, D Kleanthous, C

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Publication Type:: Journal Article
Citation:: Journal of Molecular Biology, 2015, 427 (17), pp. 2852 - 2866
Issue Date:: 2015-08-17

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Field	Value	Language
dc.contributor.author	Joshi, A	en_US
dc.contributor.author	Grinter, R	en_US
dc.contributor.author	Josts, I	en_US
dc.contributor.author	Chen, S	en_US
dc.contributor.author	Wojdyla, JA	en_US
dc.contributor.author	Lowe, ED	en_US
dc.contributor.author	Kaminska, R	en_US
dc.contributor.author	Sharp, C	en_US
dc.contributor.author	McCaughey, L https://orcid.org/0000-0001-5897-0163	en_US
dc.contributor.author	Roszak, AW	en_US
dc.contributor.author	Cogdell, RJ	en_US
dc.contributor.author	Byron, O	en_US
dc.contributor.author	Walker, D	en_US
dc.contributor.author	Kleanthous, C	en_US
dc.date.available	2015-07-20	en_US
dc.date.issued	2015-08-17	en_US
dc.identifier.citation	Journal of Molecular Biology, 2015, 427 (17), pp. 2852 - 2866	en_US
dc.identifier.issn	0022-2836	en_US
dc.identifier.uri	http://hdl.handle.net/10453/65274
dc.description.abstract	© 2015 The Authors. Published by Elsevier Ltd. How ultra-high-affinity protein-protein interactions retain high specificity is still poorly understood. The interaction between colicin DNase domains and their inhibitory immunity (Im) proteins is an ultra-high-affinity interaction that is essential for the neutralisation of endogenous DNase catalytic activity and for protection against exogenous DNase bacteriocins. The colicin DNase-Im interaction is a model system for the study of high-affinity protein-protein interactions. However, despite the fact that closely related colicin-like bacteriocins are widely produced by Gram-negative bacteria, this interaction has only been studied using colicins from Escherichia coli. In this work, we present the first crystal structures of two pyocin DNase-Im complexes from Pseudomonas aeruginosa, pyocin S2 DNase-ImS2 and pyocin AP41 DNase-ImAP41. These structures represent divergent DNase-Im subfamilies and are important in extending our understanding of protein-protein interactions for this important class of high-affinity protein complex. A key finding of this work is that mutations within the immunity protein binding energy hotspot, helix III, are tolerated by complementary substitutions at the DNase-Immunity protein binding interface. Im helix III is strictly conserved in colicins where an Asp forms polar interactions with the DNase backbone. ImAP41 contains an Asp-to-Gly substitution in helix III and our structures show the role of a co-evolved substitution where Pro in DNase loop 4 occupies the volume vacated and removes the unfulfilled hydrogen bond. We observe the co-evolved mutations in other DNase-Immunity pairs that appear to underpin the split of this family into two distinct groups.	en_US
dc.relation.ispartof	Journal of Molecular Biology	en_US
dc.relation.isbasedon	10.1016/j.jmb.2015.07.014	en_US
dc.subject.classification	Biochemistry & Molecular Biology	en_US
dc.subject.mesh	Pseudomonas aeruginosa	en_US
dc.subject.mesh	Multiprotein Complexes	en_US
dc.subject.mesh	Deoxyribonucleases	en_US
dc.subject.mesh	Pyocins	en_US
dc.subject.mesh	Colicins	en_US
dc.subject.mesh	Crystallography, X-Ray	en_US
dc.subject.mesh	Sequence Analysis, DNA	en_US
dc.subject.mesh	Protein Interaction Mapping	en_US
dc.subject.mesh	Phylogeny	en_US
dc.subject.mesh	Amino Acid Sequence	en_US
dc.subject.mesh	Base Sequence	en_US
dc.subject.mesh	Protein Structure, Tertiary	en_US
dc.subject.mesh	Protein Binding	en_US
dc.subject.mesh	Mutation	en_US
dc.subject.mesh	Molecular Sequence Data	en_US
dc.title	Structures of the Ultra-High-Affinity Protein-Protein Complexes of Pyocins S2 and AP41 and Their Cognate Immunity Proteins from Pseudomonas aeruginosa	en_US
dc.type	Journal Article
utslib.citation.volume	17	en_US
utslib.citation.volume	427	en_US
utslib.for	0601 Biochemistry and Cell Biology	en_US
utslib.for	0304 Medicinal and Biomolecular Chemistry	en_US
utslib.for	0605 Microbiology	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
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
utslib.copyright.status	open_access
pubs.issue	17	en_US
pubs.publication-status	Published	en_US
pubs.volume	427	en_US

Abstract:

© 2015 The Authors. Published by Elsevier Ltd. How ultra-high-affinity protein-protein interactions retain high specificity is still poorly understood. The interaction between colicin DNase domains and their inhibitory immunity (Im) proteins is an ultra-high-affinity interaction that is essential for the neutralisation of endogenous DNase catalytic activity and for protection against exogenous DNase bacteriocins. The colicin DNase-Im interaction is a model system for the study of high-affinity protein-protein interactions. However, despite the fact that closely related colicin-like bacteriocins are widely produced by Gram-negative bacteria, this interaction has only been studied using colicins from Escherichia coli. In this work, we present the first crystal structures of two pyocin DNase-Im complexes from Pseudomonas aeruginosa, pyocin S2 DNase-ImS2 and pyocin AP41 DNase-ImAP41. These structures represent divergent DNase-Im subfamilies and are important in extending our understanding of protein-protein interactions for this important class of high-affinity protein complex. A key finding of this work is that mutations within the immunity protein binding energy hotspot, helix III, are tolerated by complementary substitutions at the DNase-Immunity protein binding interface. Im helix III is strictly conserved in colicins where an Asp forms polar interactions with the DNase backbone. ImAP41 contains an Asp-to-Gly substitution in helix III and our structures show the role of a co-evolved substitution where Pro in DNase loop 4 occupies the volume vacated and removes the unfulfilled hydrogen bond. We observe the co-evolved mutations in other DNase-Immunity pairs that appear to underpin the split of this family into two distinct groups.

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http://hdl.handle.net/10453/65274