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Metal-dependent allosteric activation and inhibition on the same molecular scaffold: the copper sensor CopY from Streptococcus pneumoniae
Resistance to copper (Cu) toxicity in the respiratory pathogen is regulated by the Cu-specific metallosensor CopY. CopY is structurally related to the antibiotic-resistance regulatory proteins MecI and BlaI from , but is otherwise poorly characterized. Here we employ a multi-pronged experimental str...
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Published in: | Chemical science (Cambridge) 2018-01, Vol.9 (1), p.105-118 |
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creator | Glauninger, Hendrik Zhang, Yifan Higgins, Khadine A Jacobs, Alexander D Martin, Julia E Fu, Yue Coyne Rd, H Jerome Bruce, Kevin E Maroney, Michael J Clemmer, David E Capdevila, Daiana A Giedroc, David P |
description | Resistance to copper (Cu) toxicity in the respiratory pathogen
is regulated by the Cu-specific metallosensor CopY. CopY is structurally related to the antibiotic-resistance regulatory proteins MecI and BlaI from
, but is otherwise poorly characterized. Here we employ a multi-pronged experimental strategy to define the
CopY coordination chemistry and the unique mechanism of allosteric activation by Zn(ii) and allosteric inhibition by Cu(i) of
promoter DNA binding. We show that Zn(ii) is coordinated by a subunit-bridging 3S 1H
O complex formed by the same residues that coordinate Cu(i), as determined by X-ray absorption spectroscopy and ratiometric pulsed alkylation-mass spectrometry (rPA-MS). Apo- and Zn-bound CopY are homodimers by small angle X-ray scattering (SAXS); however, Zn stabilizes the dimer, narrows the conformational ensemble of the apo-state as revealed by ion mobility-mass spectroscopy (IM-MS), and activates DNA binding
and in cells. In contrast, Cu(i) employs the same Cys pair to form a subunit-bridging, kinetically stable, multi-metallic Cu·S cluster (
≈ 10
M
) that induces oligomerization beyond the dimer as revealed by SAXS, rPA-MS and NMR spectroscopy, leading to inhibition of DNA binding. These studies suggest that CopY employs conformational selection to drive Zn-activation of DNA binding, and a novel Cu(i)-mediated assembly mechanism that dissociates CopY from the DNA
ligand exchange-catalyzed metal substitution, leading to expression of Cu resistance genes. Mechanistic parallels to antibiotic resistance repressors MecI and BlaI are discussed. |
doi_str_mv | 10.1039/c7sc04396a |
format | article |
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is regulated by the Cu-specific metallosensor CopY. CopY is structurally related to the antibiotic-resistance regulatory proteins MecI and BlaI from
, but is otherwise poorly characterized. Here we employ a multi-pronged experimental strategy to define the
CopY coordination chemistry and the unique mechanism of allosteric activation by Zn(ii) and allosteric inhibition by Cu(i) of
promoter DNA binding. We show that Zn(ii) is coordinated by a subunit-bridging 3S 1H
O complex formed by the same residues that coordinate Cu(i), as determined by X-ray absorption spectroscopy and ratiometric pulsed alkylation-mass spectrometry (rPA-MS). Apo- and Zn-bound CopY are homodimers by small angle X-ray scattering (SAXS); however, Zn stabilizes the dimer, narrows the conformational ensemble of the apo-state as revealed by ion mobility-mass spectroscopy (IM-MS), and activates DNA binding
and in cells. In contrast, Cu(i) employs the same Cys pair to form a subunit-bridging, kinetically stable, multi-metallic Cu·S cluster (
≈ 10
M
) that induces oligomerization beyond the dimer as revealed by SAXS, rPA-MS and NMR spectroscopy, leading to inhibition of DNA binding. These studies suggest that CopY employs conformational selection to drive Zn-activation of DNA binding, and a novel Cu(i)-mediated assembly mechanism that dissociates CopY from the DNA
ligand exchange-catalyzed metal substitution, leading to expression of Cu resistance genes. Mechanistic parallels to antibiotic resistance repressors MecI and BlaI are discussed.</description><identifier>ISSN: 2041-6520</identifier><identifier>EISSN: 2041-6539</identifier><identifier>DOI: 10.1039/c7sc04396a</identifier><identifier>PMID: 29399317</identifier><language>eng</language><publisher>England: Royal Society of Chemistry</publisher><subject>Activation ; Alkylation ; Antibiotics ; Binding ; Chemistry ; Copper ; Deoxyribonucleic acid ; DNA ; Gene expression ; Ionic mobility ; Mass spectrometry ; NMR spectroscopy ; Oligomerization ; Proteins ; Small angle X ray scattering ; Streptococcus infections ; Toxicity</subject><ispartof>Chemical science (Cambridge), 2018-01, Vol.9 (1), p.105-118</ispartof><rights>Copyright Royal Society of Chemistry 2018</rights><rights>This journal is © The Royal Society of Chemistry 2018 2018</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c474t-afc2892370aec54075c3b831d65d1abfeca303761ec9ec023dfeb759b96dcc2f3</citedby><cites>FETCH-LOGICAL-c474t-afc2892370aec54075c3b831d65d1abfeca303761ec9ec023dfeb759b96dcc2f3</cites><orcidid>0000-0002-2342-1620 ; 0000000223421620</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5772342/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5772342/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29399317$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://www.osti.gov/biblio/1502250$$D View this record in Osti.gov$$Hfree_for_read</backlink></links><search><creatorcontrib>Glauninger, Hendrik</creatorcontrib><creatorcontrib>Zhang, Yifan</creatorcontrib><creatorcontrib>Higgins, Khadine A</creatorcontrib><creatorcontrib>Jacobs, Alexander D</creatorcontrib><creatorcontrib>Martin, Julia E</creatorcontrib><creatorcontrib>Fu, Yue</creatorcontrib><creatorcontrib>Coyne Rd, H Jerome</creatorcontrib><creatorcontrib>Bruce, Kevin E</creatorcontrib><creatorcontrib>Maroney, Michael J</creatorcontrib><creatorcontrib>Clemmer, David E</creatorcontrib><creatorcontrib>Capdevila, Daiana A</creatorcontrib><creatorcontrib>Giedroc, David P</creatorcontrib><creatorcontrib>Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)</creatorcontrib><title>Metal-dependent allosteric activation and inhibition on the same molecular scaffold: the copper sensor CopY from Streptococcus pneumoniae</title><title>Chemical science (Cambridge)</title><addtitle>Chem Sci</addtitle><description>Resistance to copper (Cu) toxicity in the respiratory pathogen
is regulated by the Cu-specific metallosensor CopY. CopY is structurally related to the antibiotic-resistance regulatory proteins MecI and BlaI from
, but is otherwise poorly characterized. Here we employ a multi-pronged experimental strategy to define the
CopY coordination chemistry and the unique mechanism of allosteric activation by Zn(ii) and allosteric inhibition by Cu(i) of
promoter DNA binding. We show that Zn(ii) is coordinated by a subunit-bridging 3S 1H
O complex formed by the same residues that coordinate Cu(i), as determined by X-ray absorption spectroscopy and ratiometric pulsed alkylation-mass spectrometry (rPA-MS). Apo- and Zn-bound CopY are homodimers by small angle X-ray scattering (SAXS); however, Zn stabilizes the dimer, narrows the conformational ensemble of the apo-state as revealed by ion mobility-mass spectroscopy (IM-MS), and activates DNA binding
and in cells. In contrast, Cu(i) employs the same Cys pair to form a subunit-bridging, kinetically stable, multi-metallic Cu·S cluster (
≈ 10
M
) that induces oligomerization beyond the dimer as revealed by SAXS, rPA-MS and NMR spectroscopy, leading to inhibition of DNA binding. These studies suggest that CopY employs conformational selection to drive Zn-activation of DNA binding, and a novel Cu(i)-mediated assembly mechanism that dissociates CopY from the DNA
ligand exchange-catalyzed metal substitution, leading to expression of Cu resistance genes. Mechanistic parallels to antibiotic resistance repressors MecI and BlaI are discussed.</description><subject>Activation</subject><subject>Alkylation</subject><subject>Antibiotics</subject><subject>Binding</subject><subject>Chemistry</subject><subject>Copper</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>Gene expression</subject><subject>Ionic mobility</subject><subject>Mass spectrometry</subject><subject>NMR spectroscopy</subject><subject>Oligomerization</subject><subject>Proteins</subject><subject>Small angle X ray scattering</subject><subject>Streptococcus infections</subject><subject>Toxicity</subject><issn>2041-6520</issn><issn>2041-6539</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><recordid>eNpdkt1qFTEQxxdRbKm98QEk6I0IR_Ox2Zx4IZTFL6h4Ub3wKmRnZz0p2WRNsgVfwafwWXwy0556UEMgk8yPf_KfTNM8ZPQ5o0K_AJWBtkJ39k5zzGnLNp0U-u4h5vSoOc35ktYhBJNc3W-OuBZaC6aOmx8fsFi_GXHBMGIoxHofc8HkgFgo7soWFwOxYSQu7NzgbrZ1lh2SbGckc_QIq7eJZLDTFP348jr56yfEZcF6iiHHRPq4fCFTijO5KAmXEiECrJksAdc5BmfxQXNvsj7j6e160nx-8_pT_25z_vHt-_7sfAOtasvGTsC3mgtFLYJsqZIghq1gYydHZocJwQoqVMcQNALlYpxwUFIPuhsB-CROmld73WUdZhyhuk7WmyW52abvJlpn_s0EtzNf45WRSnHR8irweC9QC-VMBlcQdhBDQCiGScq5pBV6entLit9WzMXMLgN6bwPGNRumdSs6yQSr6JP_0Mu4plBrYDhltJplclupZ3sKUsw54XR4MaPmuhVMry76m1Y4q_Cjvz0e0D8fL34DR0azDg</recordid><startdate>20180107</startdate><enddate>20180107</enddate><creator>Glauninger, Hendrik</creator><creator>Zhang, Yifan</creator><creator>Higgins, Khadine A</creator><creator>Jacobs, Alexander D</creator><creator>Martin, Julia E</creator><creator>Fu, Yue</creator><creator>Coyne Rd, H Jerome</creator><creator>Bruce, Kevin E</creator><creator>Maroney, Michael J</creator><creator>Clemmer, David E</creator><creator>Capdevila, Daiana A</creator><creator>Giedroc, David P</creator><general>Royal Society of Chemistry</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7SR</scope><scope>8BQ</scope><scope>8FD</scope><scope>JG9</scope><scope>7X8</scope><scope>OTOTI</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0002-2342-1620</orcidid><orcidid>https://orcid.org/0000000223421620</orcidid></search><sort><creationdate>20180107</creationdate><title>Metal-dependent allosteric activation and inhibition on the same molecular scaffold: the copper sensor CopY from Streptococcus pneumoniae</title><author>Glauninger, Hendrik ; Zhang, Yifan ; Higgins, Khadine A ; Jacobs, Alexander D ; Martin, Julia E ; Fu, Yue ; Coyne Rd, H Jerome ; Bruce, Kevin E ; Maroney, Michael J ; Clemmer, David E ; Capdevila, Daiana A ; Giedroc, David P</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c474t-afc2892370aec54075c3b831d65d1abfeca303761ec9ec023dfeb759b96dcc2f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Activation</topic><topic>Alkylation</topic><topic>Antibiotics</topic><topic>Binding</topic><topic>Chemistry</topic><topic>Copper</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>Gene expression</topic><topic>Ionic mobility</topic><topic>Mass spectrometry</topic><topic>NMR spectroscopy</topic><topic>Oligomerization</topic><topic>Proteins</topic><topic>Small angle X ray scattering</topic><topic>Streptococcus infections</topic><topic>Toxicity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Glauninger, Hendrik</creatorcontrib><creatorcontrib>Zhang, Yifan</creatorcontrib><creatorcontrib>Higgins, Khadine A</creatorcontrib><creatorcontrib>Jacobs, Alexander D</creatorcontrib><creatorcontrib>Martin, Julia E</creatorcontrib><creatorcontrib>Fu, Yue</creatorcontrib><creatorcontrib>Coyne Rd, H Jerome</creatorcontrib><creatorcontrib>Bruce, Kevin E</creatorcontrib><creatorcontrib>Maroney, Michael J</creatorcontrib><creatorcontrib>Clemmer, David E</creatorcontrib><creatorcontrib>Capdevila, Daiana A</creatorcontrib><creatorcontrib>Giedroc, David P</creatorcontrib><creatorcontrib>Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>Engineered Materials Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Materials Research Database</collection><collection>MEDLINE - Academic</collection><collection>OSTI.GOV</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Chemical science (Cambridge)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Glauninger, Hendrik</au><au>Zhang, Yifan</au><au>Higgins, Khadine A</au><au>Jacobs, Alexander D</au><au>Martin, Julia E</au><au>Fu, Yue</au><au>Coyne Rd, H Jerome</au><au>Bruce, Kevin E</au><au>Maroney, Michael J</au><au>Clemmer, David E</au><au>Capdevila, Daiana A</au><au>Giedroc, David P</au><aucorp>Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)</aucorp><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Metal-dependent allosteric activation and inhibition on the same molecular scaffold: the copper sensor CopY from Streptococcus pneumoniae</atitle><jtitle>Chemical science (Cambridge)</jtitle><addtitle>Chem Sci</addtitle><date>2018-01-07</date><risdate>2018</risdate><volume>9</volume><issue>1</issue><spage>105</spage><epage>118</epage><pages>105-118</pages><issn>2041-6520</issn><eissn>2041-6539</eissn><abstract>Resistance to copper (Cu) toxicity in the respiratory pathogen
is regulated by the Cu-specific metallosensor CopY. CopY is structurally related to the antibiotic-resistance regulatory proteins MecI and BlaI from
, but is otherwise poorly characterized. Here we employ a multi-pronged experimental strategy to define the
CopY coordination chemistry and the unique mechanism of allosteric activation by Zn(ii) and allosteric inhibition by Cu(i) of
promoter DNA binding. We show that Zn(ii) is coordinated by a subunit-bridging 3S 1H
O complex formed by the same residues that coordinate Cu(i), as determined by X-ray absorption spectroscopy and ratiometric pulsed alkylation-mass spectrometry (rPA-MS). Apo- and Zn-bound CopY are homodimers by small angle X-ray scattering (SAXS); however, Zn stabilizes the dimer, narrows the conformational ensemble of the apo-state as revealed by ion mobility-mass spectroscopy (IM-MS), and activates DNA binding
and in cells. In contrast, Cu(i) employs the same Cys pair to form a subunit-bridging, kinetically stable, multi-metallic Cu·S cluster (
≈ 10
M
) that induces oligomerization beyond the dimer as revealed by SAXS, rPA-MS and NMR spectroscopy, leading to inhibition of DNA binding. These studies suggest that CopY employs conformational selection to drive Zn-activation of DNA binding, and a novel Cu(i)-mediated assembly mechanism that dissociates CopY from the DNA
ligand exchange-catalyzed metal substitution, leading to expression of Cu resistance genes. Mechanistic parallels to antibiotic resistance repressors MecI and BlaI are discussed.</abstract><cop>England</cop><pub>Royal Society of Chemistry</pub><pmid>29399317</pmid><doi>10.1039/c7sc04396a</doi><tpages>14</tpages><orcidid>https://orcid.org/0000-0002-2342-1620</orcidid><orcidid>https://orcid.org/0000000223421620</orcidid><oa>free_for_read</oa></addata></record> |
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subjects | Activation Alkylation Antibiotics Binding Chemistry Copper Deoxyribonucleic acid DNA Gene expression Ionic mobility Mass spectrometry NMR spectroscopy Oligomerization Proteins Small angle X ray scattering Streptococcus infections Toxicity |
title | Metal-dependent allosteric activation and inhibition on the same molecular scaffold: the copper sensor CopY from Streptococcus pneumoniae |
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