Methylerythritol Phosphate Pathway: Enzymatic Evidence for a Rotation in the LytB/IspH‐Catalyzed Reaction
IspH/LytB, an oxygen‐sensitive [4Fe‐4S] enzyme, catalyzes the last step of the methylerythritol phosphate (MEP) pathway, a target for the development of new antimicrobial agents. This metalloenzyme converts (E)‐4‐hydroxy‐3‐methylbut‐2‐en‐1‐yl diphosphate (HMBPP) into the two isoprenoid precursors: i...
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| Veröffentlicht in: | Chemistry : a European journal 2020-01, Vol.26 (5), p.1032-1036 |
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| creator | Chaignon, Philippe Petit, Benoît Eric Vincent, Bruno Allouche, Lionel Seemann, Myriam |
| description | IspH/LytB, an oxygen‐sensitive [4Fe‐4S] enzyme, catalyzes the last step of the methylerythritol phosphate (MEP) pathway, a target for the development of new antimicrobial agents. This metalloenzyme converts (E)‐4‐hydroxy‐3‐methylbut‐2‐en‐1‐yl diphosphate (HMBPP) into the two isoprenoid precursors: isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). Here, the synthesis of (S)‐[4‐2H1]HMBPP and (R)‐[4‐2H1]HMBPP is reported together with a detailed NMR analysis of the products formed after their respective incubation with E. coli IspH/LytB in the presence of the biological reduction system used by E. coli to reduce the [4Fe‐4S] center. (S)‐[4‐2H1]HMBPP was converted into [4‐2H1]DMAPP and (E)‐[4‐2H1]IPP, whereas (R)‐[4‐2H1]HMBPP yielded [4‐2H1]DMAPP and (Z)‐[4‐2H1]IPP, hence providing the direct enzymatic evidence that the mechanism catalyzed by IspH/LytB involves a rotation of the CH2OH group of the substrate to display it away from the [4Fe‐4S].
A reductive dehydroxylation is catalyzed by IspH/LytB, a [4Fe‐4S] enzyme target for the development of innovative antibacterial agents. The missing enzymatic evidence in the investigation of a rotation of the CH2OH group of the substrate in the catalytic mechanism of this enzyme is provided after synthesis of deuterium‐labeled stereoisomers of the substrate followed by the analysis of the products by several NMR methodologies. |
| doi_str_mv | 10.1002/chem.201904676 |
| format | Article |
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A reductive dehydroxylation is catalyzed by IspH/LytB, a [4Fe‐4S] enzyme target for the development of innovative antibacterial agents. The missing enzymatic evidence in the investigation of a rotation of the CH2OH group of the substrate in the catalytic mechanism of this enzyme is provided after synthesis of deuterium‐labeled stereoisomers of the substrate followed by the analysis of the products by several NMR methodologies.</description><identifier>ISSN: 0947-6539</identifier><identifier>EISSN: 1521-3765</identifier><identifier>DOI: 10.1002/chem.201904676</identifier><identifier>PMID: 31756006</identifier><language>eng</language><publisher>Germany: Wiley Subscription Services, Inc</publisher><subject>[4Fe-4S] cluster ; Antimicrobial agents ; Biocatalysis ; Catalysis ; Chemical Sciences ; Chemistry ; E coli ; enzyme catalysis ; Escherichia coli - enzymology ; Escherichia coli Proteins - metabolism ; Isopentenyl diphosphate ; IspH/LytB ; Life Sciences ; metalloenzymes ; methylerythritol phosphate pathway ; NMR ; Nuclear magnetic resonance ; Organic chemistry ; Organophosphates - chemistry ; Organophosphates - metabolism ; Oxidation-Reduction ; Oxidoreductases - metabolism ; Phosphates - chemistry ; Phosphates - metabolism ; Rotation ; Substrate Specificity ; Substrates ; Terpenes - chemistry ; Terpenes - metabolism</subject><ispartof>Chemistry : a European journal, 2020-01, Vol.26 (5), p.1032-1036</ispartof><rights>2019 Wiley‐VCH Verlag GmbH & Co. KGaA, Weinheim</rights><rights>2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.</rights><rights>2020 Wiley‐VCH Verlag GmbH & Co. KGaA, Weinheim</rights><rights>Distributed under a Creative Commons Attribution 4.0 International License</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4846-8410bdd935b2ec97f6c89172a0df7620f8740ab95bd2dcde1c7440957862d53c3</citedby><cites>FETCH-LOGICAL-c4846-8410bdd935b2ec97f6c89172a0df7620f8740ab95bd2dcde1c7440957862d53c3</cites><orcidid>0000-0002-4766-3958 ; 0000-0002-2615-1574</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fchem.201904676$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fchem.201904676$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>230,314,780,784,885,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31756006$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://hal.science/hal-02407672$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Chaignon, Philippe</creatorcontrib><creatorcontrib>Petit, Benoît Eric</creatorcontrib><creatorcontrib>Vincent, Bruno</creatorcontrib><creatorcontrib>Allouche, Lionel</creatorcontrib><creatorcontrib>Seemann, Myriam</creatorcontrib><title>Methylerythritol Phosphate Pathway: Enzymatic Evidence for a Rotation in the LytB/IspH‐Catalyzed Reaction</title><title>Chemistry : a European journal</title><addtitle>Chemistry</addtitle><description>IspH/LytB, an oxygen‐sensitive [4Fe‐4S] enzyme, catalyzes the last step of the methylerythritol phosphate (MEP) pathway, a target for the development of new antimicrobial agents. This metalloenzyme converts (E)‐4‐hydroxy‐3‐methylbut‐2‐en‐1‐yl diphosphate (HMBPP) into the two isoprenoid precursors: isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). Here, the synthesis of (S)‐[4‐2H1]HMBPP and (R)‐[4‐2H1]HMBPP is reported together with a detailed NMR analysis of the products formed after their respective incubation with E. coli IspH/LytB in the presence of the biological reduction system used by E. coli to reduce the [4Fe‐4S] center. (S)‐[4‐2H1]HMBPP was converted into [4‐2H1]DMAPP and (E)‐[4‐2H1]IPP, whereas (R)‐[4‐2H1]HMBPP yielded [4‐2H1]DMAPP and (Z)‐[4‐2H1]IPP, hence providing the direct enzymatic evidence that the mechanism catalyzed by IspH/LytB involves a rotation of the CH2OH group of the substrate to display it away from the [4Fe‐4S].
A reductive dehydroxylation is catalyzed by IspH/LytB, a [4Fe‐4S] enzyme target for the development of innovative antibacterial agents. The missing enzymatic evidence in the investigation of a rotation of the CH2OH group of the substrate in the catalytic mechanism of this enzyme is provided after synthesis of deuterium‐labeled stereoisomers of the substrate followed by the analysis of the products by several NMR methodologies.</description><subject>[4Fe-4S] cluster</subject><subject>Antimicrobial agents</subject><subject>Biocatalysis</subject><subject>Catalysis</subject><subject>Chemical Sciences</subject><subject>Chemistry</subject><subject>E coli</subject><subject>enzyme catalysis</subject><subject>Escherichia coli - enzymology</subject><subject>Escherichia coli Proteins - metabolism</subject><subject>Isopentenyl diphosphate</subject><subject>IspH/LytB</subject><subject>Life Sciences</subject><subject>metalloenzymes</subject><subject>methylerythritol phosphate pathway</subject><subject>NMR</subject><subject>Nuclear magnetic resonance</subject><subject>Organic chemistry</subject><subject>Organophosphates - chemistry</subject><subject>Organophosphates - metabolism</subject><subject>Oxidation-Reduction</subject><subject>Oxidoreductases - metabolism</subject><subject>Phosphates - chemistry</subject><subject>Phosphates - metabolism</subject><subject>Rotation</subject><subject>Substrate Specificity</subject><subject>Substrates</subject><subject>Terpenes - chemistry</subject><subject>Terpenes - metabolism</subject><issn>0947-6539</issn><issn>1521-3765</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0UFv0zAYBmALgVgZXDkiS1zgkO6z49gxt60qdFInpgnOlmM7SkYSl9jdlJ34CfxGfgmOOorEhZOlT49f2d-L0GsCSwJAz0zj-iUFIoFxwZ-gBSkoyXLBi6doAZKJjBe5PEEvQrgFAMnz_Dk6yYkoOABfoG9XLjZT58YpNmMbfYevGx92jY4OX-vY3OvpA14PD1OvY2vw-q61bjAO137EGt_4mMZ-wO2AY-PwdooXZ5dht_n14-dKR91ND87iG6fNrF6iZ7Xugnv1eJ6irx_XX1abbPv50-XqfJsZVjKelYxAZa3Mi4o6I0XNTSmJoBpsLTiFuhQMdCWLylJrrCNGMAayECWntshNforeH3Ib3and2PZ6nJTXrdqcb9U8A8pAcEHvSLLvDnY3-u97F6Lq22Bc1-nB-X1QNK2KA2GCJvr2H3rr9-OQfpJUAhJEMavlQZnRhzC6-vgCAmquTM2VqWNl6cKbx9h91Tt75H86SkAewH3buek_cWq1WV_9Df8N2vWigw</recordid><startdate>20200122</startdate><enddate>20200122</enddate><creator>Chaignon, Philippe</creator><creator>Petit, Benoît Eric</creator><creator>Vincent, Bruno</creator><creator>Allouche, Lionel</creator><creator>Seemann, Myriam</creator><general>Wiley Subscription Services, Inc</general><general>Wiley-VCH Verlag</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7SR</scope><scope>8BQ</scope><scope>8FD</scope><scope>JG9</scope><scope>K9.</scope><scope>7X8</scope><scope>1XC</scope><scope>VOOES</scope><orcidid>https://orcid.org/0000-0002-4766-3958</orcidid><orcidid>https://orcid.org/0000-0002-2615-1574</orcidid></search><sort><creationdate>20200122</creationdate><title>Methylerythritol Phosphate Pathway: Enzymatic Evidence for a Rotation in the LytB/IspH‐Catalyzed Reaction</title><author>Chaignon, Philippe ; Petit, Benoît Eric ; Vincent, Bruno ; Allouche, Lionel ; Seemann, Myriam</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4846-8410bdd935b2ec97f6c89172a0df7620f8740ab95bd2dcde1c7440957862d53c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>[4Fe-4S] cluster</topic><topic>Antimicrobial agents</topic><topic>Biocatalysis</topic><topic>Catalysis</topic><topic>Chemical Sciences</topic><topic>Chemistry</topic><topic>E coli</topic><topic>enzyme catalysis</topic><topic>Escherichia coli - enzymology</topic><topic>Escherichia coli Proteins - metabolism</topic><topic>Isopentenyl diphosphate</topic><topic>IspH/LytB</topic><topic>Life Sciences</topic><topic>metalloenzymes</topic><topic>methylerythritol phosphate pathway</topic><topic>NMR</topic><topic>Nuclear magnetic resonance</topic><topic>Organic chemistry</topic><topic>Organophosphates - chemistry</topic><topic>Organophosphates - metabolism</topic><topic>Oxidation-Reduction</topic><topic>Oxidoreductases - metabolism</topic><topic>Phosphates - chemistry</topic><topic>Phosphates - metabolism</topic><topic>Rotation</topic><topic>Substrate Specificity</topic><topic>Substrates</topic><topic>Terpenes - chemistry</topic><topic>Terpenes - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chaignon, Philippe</creatorcontrib><creatorcontrib>Petit, Benoît Eric</creatorcontrib><creatorcontrib>Vincent, Bruno</creatorcontrib><creatorcontrib>Allouche, Lionel</creatorcontrib><creatorcontrib>Seemann, Myriam</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><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>ProQuest Health & Medical Complete (Alumni)</collection><collection>MEDLINE - Academic</collection><collection>Hyper Article en Ligne (HAL)</collection><collection>Hyper Article en Ligne (HAL) (Open Access)</collection><jtitle>Chemistry : a European journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chaignon, Philippe</au><au>Petit, Benoît Eric</au><au>Vincent, Bruno</au><au>Allouche, Lionel</au><au>Seemann, Myriam</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Methylerythritol Phosphate Pathway: Enzymatic Evidence for a Rotation in the LytB/IspH‐Catalyzed Reaction</atitle><jtitle>Chemistry : a European journal</jtitle><addtitle>Chemistry</addtitle><date>2020-01-22</date><risdate>2020</risdate><volume>26</volume><issue>5</issue><spage>1032</spage><epage>1036</epage><pages>1032-1036</pages><issn>0947-6539</issn><eissn>1521-3765</eissn><abstract>IspH/LytB, an oxygen‐sensitive [4Fe‐4S] enzyme, catalyzes the last step of the methylerythritol phosphate (MEP) pathway, a target for the development of new antimicrobial agents. This metalloenzyme converts (E)‐4‐hydroxy‐3‐methylbut‐2‐en‐1‐yl diphosphate (HMBPP) into the two isoprenoid precursors: isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). Here, the synthesis of (S)‐[4‐2H1]HMBPP and (R)‐[4‐2H1]HMBPP is reported together with a detailed NMR analysis of the products formed after their respective incubation with E. coli IspH/LytB in the presence of the biological reduction system used by E. coli to reduce the [4Fe‐4S] center. (S)‐[4‐2H1]HMBPP was converted into [4‐2H1]DMAPP and (E)‐[4‐2H1]IPP, whereas (R)‐[4‐2H1]HMBPP yielded [4‐2H1]DMAPP and (Z)‐[4‐2H1]IPP, hence providing the direct enzymatic evidence that the mechanism catalyzed by IspH/LytB involves a rotation of the CH2OH group of the substrate to display it away from the [4Fe‐4S].
A reductive dehydroxylation is catalyzed by IspH/LytB, a [4Fe‐4S] enzyme target for the development of innovative antibacterial agents. The missing enzymatic evidence in the investigation of a rotation of the CH2OH group of the substrate in the catalytic mechanism of this enzyme is provided after synthesis of deuterium‐labeled stereoisomers of the substrate followed by the analysis of the products by several NMR methodologies.</abstract><cop>Germany</cop><pub>Wiley Subscription Services, Inc</pub><pmid>31756006</pmid><doi>10.1002/chem.201904676</doi><tpages>5</tpages><orcidid>https://orcid.org/0000-0002-4766-3958</orcidid><orcidid>https://orcid.org/0000-0002-2615-1574</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | [4Fe-4S] cluster Antimicrobial agents Biocatalysis Catalysis Chemical Sciences Chemistry E coli enzyme catalysis Escherichia coli - enzymology Escherichia coli Proteins - metabolism Isopentenyl diphosphate IspH/LytB Life Sciences metalloenzymes methylerythritol phosphate pathway NMR Nuclear magnetic resonance Organic chemistry Organophosphates - chemistry Organophosphates - metabolism Oxidation-Reduction Oxidoreductases - metabolism Phosphates - chemistry Phosphates - metabolism Rotation Substrate Specificity Substrates Terpenes - chemistry Terpenes - metabolism |
| title | Methylerythritol Phosphate Pathway: Enzymatic Evidence for a Rotation in the LytB/IspH‐Catalyzed Reaction |
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