Single-Molecule Force Spectroscopy Reveals that the Fe-N Bond Enables Multiple Rupture Pathways of the 2Fe2S Cluster in a MitoNEET Monomer.
Identifieur interne : 000043 ( Main/Exploration ); précédent : 000042; suivant : 000044Single-Molecule Force Spectroscopy Reveals that the Fe-N Bond Enables Multiple Rupture Pathways of the 2Fe2S Cluster in a MitoNEET Monomer.
Auteurs : Guobin Song [République populaire de Chine] ; Xuan Ding [République populaire de Chine] ; Huaxing Liu [République populaire de Chine] ; Guodong Yuan [République populaire de Chine] ; Fang Tian [République populaire de Chine] ; Shengchao Shi [République populaire de Chine] ; Yang Yang [République populaire de Chine] ; Guoqiang Li [République populaire de Chine] ; Peng Zheng [République populaire de Chine]Source :
- Analytical chemistry [ 1520-6882 ] ; 2020.
Abstract
The mitochondrial outer membrane protein, mitoNEET (mNT), is an iron-sulfur protein containing an Fe2S2(His)1(Cys)3 cluster with a unique single Fe-N bond. Previous studies have shown that this Fe(III)-N(His) bond is essential for metal cluster transfer and protein function. To further understand the effect of this unique Fe-N bond on the metal cluster and protein, we used atomic force microscopy-based single-molecule force spectroscopy (AFM-SMFS) to investigate the mechanical unfolding mechanism of an mNT monomer, focusing on the rupture pathway and kinetic stability of the cluster. We found that the Fe-N bond was the weakest point of the cluster, the rupture of which occurred first, and could be independent of the cluster break. Moreover, this Fe-N bond enabled a dynamic and labile iron-sulfur cluster, as multiple unfolding pathways of mNT with a unique Fe2S2(Cys)3 intermediate were observed accordingly.
DOI: 10.1021/acs.analchem.0c03536
PubMed: 33048522
Affiliations:
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China.</nlm:affiliation><country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023</wicri:regionArea><wicri:noRegion>Jiangsu 210023</wicri:noRegion></affiliation></author><author><name sortKey="Yang, Yang" sort="Yang, Yang" uniqKey="Yang Y" first="Yang" last="Yang">Yang Yang</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</nlm:affiliation><country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023</wicri:regionArea><wicri:noRegion>Jiangsu 210023</wicri:noRegion></affiliation></author><author><name sortKey="Li, Guoqiang" sort="Li, Guoqiang" uniqKey="Li G" first="Guoqiang" last="Li">Guoqiang Li</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. 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R. China.</nlm:affiliation><country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023</wicri:regionArea><wicri:noRegion>Jiangsu 210023</wicri:noRegion></affiliation></author><author><name sortKey="Ding, Xuan" sort="Ding, Xuan" uniqKey="Ding X" first="Xuan" last="Ding">Xuan Ding</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</nlm:affiliation><country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023</wicri:regionArea><wicri:noRegion>Jiangsu 210023</wicri:noRegion></affiliation></author><author><name sortKey="Liu, Huaxing" sort="Liu, Huaxing" uniqKey="Liu H" first="Huaxing" last="Liu">Huaxing Liu</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</nlm:affiliation><country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023</wicri:regionArea><wicri:noRegion>Jiangsu 210023</wicri:noRegion></affiliation></author><author><name sortKey="Yuan, Guodong" sort="Yuan, Guodong" uniqKey="Yuan G" first="Guodong" last="Yuan">Guodong Yuan</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</nlm:affiliation><country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023</wicri:regionArea><wicri:noRegion>Jiangsu 210023</wicri:noRegion></affiliation></author><author><name sortKey="Tian, Fang" sort="Tian, Fang" uniqKey="Tian F" first="Fang" last="Tian">Fang Tian</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</nlm:affiliation><country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023</wicri:regionArea><wicri:noRegion>Jiangsu 210023</wicri:noRegion></affiliation></author><author><name sortKey="Shi, Shengchao" sort="Shi, Shengchao" uniqKey="Shi S" first="Shengchao" last="Shi">Shengchao Shi</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</nlm:affiliation><country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023</wicri:regionArea><wicri:noRegion>Jiangsu 210023</wicri:noRegion></affiliation></author><author><name sortKey="Yang, Yang" sort="Yang, Yang" uniqKey="Yang Y" first="Yang" last="Yang">Yang Yang</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</nlm:affiliation><country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023</wicri:regionArea><wicri:noRegion>Jiangsu 210023</wicri:noRegion></affiliation></author><author><name sortKey="Li, Guoqiang" sort="Li, Guoqiang" uniqKey="Li G" first="Guoqiang" last="Li">Guoqiang Li</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</nlm:affiliation><country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023</wicri:regionArea><wicri:noRegion>Jiangsu 210023</wicri:noRegion></affiliation></author><author><name sortKey="Zheng, Peng" sort="Zheng, Peng" uniqKey="Zheng P" first="Peng" last="Zheng">Peng Zheng</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</nlm:affiliation><country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023</wicri:regionArea><wicri:noRegion>Jiangsu 210023</wicri:noRegion></affiliation></author></analytic><series><title level="j">Analytical chemistry</title><idno type="eISSN">1520-6882</idno><imprint><date when="2020" type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The mitochondrial outer membrane protein, mitoNEET (mNT), is an iron-sulfur protein containing an Fe<sub>2</sub>S<sub>2</sub>(His)<sub>1</sub>(Cys)<sub>3</sub> cluster with a unique single Fe-N bond. Previous studies have shown that this Fe(III)-N(His) bond is essential for metal cluster transfer and protein function. To further understand the effect of this unique Fe-N bond on the metal cluster and protein, we used atomic force microscopy-based single-molecule force spectroscopy (AFM-SMFS) to investigate the mechanical unfolding mechanism of an mNT monomer, focusing on the rupture pathway and kinetic stability of the cluster. We found that the Fe-N bond was the weakest point of the cluster, the rupture of which occurred first, and could be independent of the cluster break. Moreover, this Fe-N bond enabled a dynamic and labile iron-sulfur cluster, as multiple unfolding pathways of mNT with a unique Fe<sub>2</sub>S<sub>2</sub>(Cys)<sub>3</sub> intermediate were observed accordingly.</div></front></TEI><pubmed><MedlineCitation Status="In-Data-Review" Owner="NLM"><PMID Version="1">33048522</PMID><DateRevised><Year>2020</Year><Month>11</Month><Day>03</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1520-6882</ISSN><JournalIssue CitedMedium="Internet"><Volume>92</Volume><Issue>21</Issue><PubDate><Year>2020</Year><Month>Nov</Month><Day>03</Day></PubDate></JournalIssue><Title>Analytical chemistry</Title><ISOAbbreviation>Anal Chem</ISOAbbreviation></Journal><ArticleTitle>Single-Molecule Force Spectroscopy Reveals that the Fe-N Bond Enables Multiple Rupture Pathways of the 2Fe2S Cluster in a MitoNEET Monomer.</ArticleTitle><Pagination><MedlinePgn>14783-14789</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1021/acs.analchem.0c03536</ELocationID><Abstract><AbstractText>The mitochondrial outer membrane protein, mitoNEET (mNT), is an iron-sulfur protein containing an Fe<sub>2</sub>S<sub>2</sub>(His)<sub>1</sub>(Cys)<sub>3</sub> cluster with a unique single Fe-N bond. Previous studies have shown that this Fe(III)-N(His) bond is essential for metal cluster transfer and protein function. To further understand the effect of this unique Fe-N bond on the metal cluster and protein, we used atomic force microscopy-based single-molecule force spectroscopy (AFM-SMFS) to investigate the mechanical unfolding mechanism of an mNT monomer, focusing on the rupture pathway and kinetic stability of the cluster. We found that the Fe-N bond was the weakest point of the cluster, the rupture of which occurred first, and could be independent of the cluster break. Moreover, this Fe-N bond enabled a dynamic and labile iron-sulfur cluster, as multiple unfolding pathways of mNT with a unique Fe<sub>2</sub>S<sub>2</sub>(Cys)<sub>3</sub> intermediate were observed accordingly.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Song</LastName><ForeName>Guobin</ForeName><Initials>G</Initials><AffiliationInfo><Affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Ding</LastName><ForeName>Xuan</ForeName><Initials>X</Initials><AffiliationInfo><Affiliation>State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Liu</LastName><ForeName>Huaxing</ForeName><Initials>H</Initials><AffiliationInfo><Affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Yuan</LastName><ForeName>Guodong</ForeName><Initials>G</Initials><AffiliationInfo><Affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Tian</LastName><ForeName>Fang</ForeName><Initials>F</Initials><AffiliationInfo><Affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Shi</LastName><ForeName>Shengchao</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Yang</LastName><ForeName>Yang</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Li</LastName><ForeName>Guoqiang</ForeName><Initials>G</Initials><AffiliationInfo><Affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zheng</LastName><ForeName>Peng</ForeName><Initials>P</Initials><AffiliationInfo><Affiliation>State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210023, P. R. China.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2020</Year><Month>10</Month><Day>13</Day></ArticleDate></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Anal Chem</MedlineTA><NlmUniqueID>0370536</NlmUniqueID><ISSNLinking>0003-2700</ISSNLinking></MedlineJournalInfo><CitationSubset>IM</CitationSubset></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2020</Year><Month>10</Month><Day>14</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2020</Year><Month>10</Month><Day>14</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2020</Year><Month>10</Month><Day>13</Day><Hour>17</Hour><Minute>9</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">33048522</ArticleId><ArticleId IdType="doi">10.1021/acs.analchem.0c03536</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>République populaire de Chine</li></country></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Song, Guobin" sort="Song, Guobin" uniqKey="Song G" first="Guobin" last="Song">Guobin Song</name></noRegion><name sortKey="Ding, Xuan" sort="Ding, Xuan" uniqKey="Ding X" first="Xuan" last="Ding">Xuan Ding</name><name sortKey="Li, Guoqiang" sort="Li, Guoqiang" uniqKey="Li G" first="Guoqiang" last="Li">Guoqiang Li</name><name sortKey="Liu, Huaxing" sort="Liu, Huaxing" uniqKey="Liu H" first="Huaxing" last="Liu">Huaxing Liu</name><name sortKey="Shi, Shengchao" sort="Shi, Shengchao" uniqKey="Shi S" first="Shengchao" last="Shi">Shengchao Shi</name><name sortKey="Tian, Fang" sort="Tian, Fang" uniqKey="Tian F" first="Fang" last="Tian">Fang Tian</name><name sortKey="Yang, Yang" sort="Yang, Yang" uniqKey="Yang Y" first="Yang" last="Yang">Yang Yang</name><name sortKey="Yuan, Guodong" sort="Yuan, Guodong" uniqKey="Yuan G" first="Guodong" last="Yuan">Guodong Yuan</name><name sortKey="Zheng, Peng" sort="Zheng, Peng" uniqKey="Zheng P" first="Peng" last="Zheng">Peng Zheng</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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