NMR Spectroscopic Characterization of the C‐Mannose Conformation in a Thrombospondin Repeat Using a Selective Labeling Approach
Despite the great interest in glycoproteins, structural information reporting on conformation and dynamics of the sugar moieties are limited. We present a new biochemical method to express proteins with glycans that are selectively labeled with NMR‐active nuclei. We report on the incorporation of 13...
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| Veröffentlicht in: | Angewandte Chemie International Edition 2020-11, Vol.59 (46), p.20659-20665 |
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| creator | Jonker, Hendrik R. A. Saxena, Krishna Shcherbakova, Aleksandra Tiemann, Birgit Bakker, Hans Schwalbe, Harald |
| description | Despite the great interest in glycoproteins, structural information reporting on conformation and dynamics of the sugar moieties are limited. We present a new biochemical method to express proteins with glycans that are selectively labeled with NMR‐active nuclei. We report on the incorporation of 13C‐labeled mannose in the C‐mannosylated UNC‐5 thrombospondin repeat. The conformational landscape of the C‐mannose sugar puckers attached to tryptophan residues of UNC‐5 is characterized by interconversion between the canonical 1C4 state and the B03 / 1S3 state. This flexibility may be essential for protein folding and stabilization. We foresee that this versatile tool to produce proteins with selectively labeled C‐mannose can be applied and adjusted to other systems and modifications and potentially paves a way to advance glycoprotein research by unravelling the dynamical and conformational properties of glycan structures and their interactions.
NMR spectroscopy was used to characterize the C‐mannose conformation in a thrombospondin repeat using a selective labeling approach. The conformational landscape of the C‐mannose sugar puckers attached to tryptophan residues of the netrin receptor UNC‐5 allows interconversion between the canonical 1C4 state and the B03 / 1S3 state. |
| doi_str_mv | 10.1002/anie.202009489 |
| format | Article |
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NMR spectroscopy was used to characterize the C‐mannose conformation in a thrombospondin repeat using a selective labeling approach. The conformational landscape of the C‐mannose sugar puckers attached to tryptophan residues of the netrin receptor UNC‐5 allows interconversion between the canonical 1C4 state and the B03 / 1S3 state.</description><edition>International ed. in English</edition><identifier>ISSN: 1433-7851</identifier><identifier>EISSN: 1521-3773</identifier><identifier>DOI: 10.1002/anie.202009489</identifier><identifier>PMID: 32745319</identifier><language>eng</language><publisher>Germany: Wiley Subscription Services, Inc</publisher><subject>C-mannosylation ; conformation analysis ; Glycan ; Glycoproteins ; Mannose ; NMR ; NMR spectroscopy ; Nuclear magnetic resonance ; Polysaccharides ; Protein folding ; Protein structure ; Proteins ; selective isotopic labeling ; Thrombospondin ; Tryptophan</subject><ispartof>Angewandte Chemie International Edition, 2020-11, Vol.59 (46), p.20659-20665</ispartof><rights>2020 The Authors. Published by Wiley-VCH GmbH</rights><rights>2020 The Authors. Published by Wiley-VCH GmbH.</rights><rights>2020. This article is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5719-67f3a9b36221b8c4a35c8e15eda1ae036e5dfd71c9beb51df58fb2e0d5a5911d3</citedby><cites>FETCH-LOGICAL-c5719-67f3a9b36221b8c4a35c8e15eda1ae036e5dfd71c9beb51df58fb2e0d5a5911d3</cites><orcidid>0000-0003-4175-547X ; 0000-0001-5693-7909 ; 0000-0002-2237-5378 ; 0000-0002-5582-3931 ; 0000-0002-1364-9154</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%2Fanie.202009489$$EPDF$$P50$$Gwiley$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fanie.202009489$$EHTML$$P50$$Gwiley$$Hfree_for_read</linktohtml><link.rule.ids>230,314,780,784,885,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32745319$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Jonker, Hendrik R. A.</creatorcontrib><creatorcontrib>Saxena, Krishna</creatorcontrib><creatorcontrib>Shcherbakova, Aleksandra</creatorcontrib><creatorcontrib>Tiemann, Birgit</creatorcontrib><creatorcontrib>Bakker, Hans</creatorcontrib><creatorcontrib>Schwalbe, Harald</creatorcontrib><title>NMR Spectroscopic Characterization of the C‐Mannose Conformation in a Thrombospondin Repeat Using a Selective Labeling Approach</title><title>Angewandte Chemie International Edition</title><addtitle>Angew Chem Int Ed Engl</addtitle><description>Despite the great interest in glycoproteins, structural information reporting on conformation and dynamics of the sugar moieties are limited. We present a new biochemical method to express proteins with glycans that are selectively labeled with NMR‐active nuclei. We report on the incorporation of 13C‐labeled mannose in the C‐mannosylated UNC‐5 thrombospondin repeat. The conformational landscape of the C‐mannose sugar puckers attached to tryptophan residues of UNC‐5 is characterized by interconversion between the canonical 1C4 state and the B03 / 1S3 state. This flexibility may be essential for protein folding and stabilization. We foresee that this versatile tool to produce proteins with selectively labeled C‐mannose can be applied and adjusted to other systems and modifications and potentially paves a way to advance glycoprotein research by unravelling the dynamical and conformational properties of glycan structures and their interactions.
NMR spectroscopy was used to characterize the C‐mannose conformation in a thrombospondin repeat using a selective labeling approach. The conformational landscape of the C‐mannose sugar puckers attached to tryptophan residues of the netrin receptor UNC‐5 allows interconversion between the canonical 1C4 state and the B03 / 1S3 state.</description><subject>C-mannosylation</subject><subject>conformation analysis</subject><subject>Glycan</subject><subject>Glycoproteins</subject><subject>Mannose</subject><subject>NMR</subject><subject>NMR spectroscopy</subject><subject>Nuclear magnetic resonance</subject><subject>Polysaccharides</subject><subject>Protein folding</subject><subject>Protein structure</subject><subject>Proteins</subject><subject>selective isotopic labeling</subject><subject>Thrombospondin</subject><subject>Tryptophan</subject><issn>1433-7851</issn><issn>1521-3773</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>24P</sourceid><sourceid>WIN</sourceid><recordid>eNqFkctu1DAUhi0Eou3AliWKxKabGXyJk3iDNBqVUmlapF7W1olz0rhK7GBnisoK3oBn5EnwaMpw2bDy0fk__8fHPyGvGF0wSvlbcBYXnHJKVV6pJ-SQSc7moizF01TnQszLSrIDchTjXeKrihbPyYHgZS4FU4fk28X5ZXY1opmCj8aP1mSrDgKYCYP9ApP1LvNtNnWYrX58_X4OzvmYau9aH4adbl0G2XUX_FD7OHrXpMYljghTdhOtu03qFfZphL3HbA019tvmchyDB9O9IM9a6CO-fDxn5Ob9yfXqw3z98fRstVzPjSyZmhdlK0DVouCc1ZXJQUhTIZPYAAOkokDZtE3JjKqxlqxpZdXWHGkjQSrGGjEj73a-46YesDHopgC9HoMdIDxoD1b_rTjb6Vt_r8tCcSVZMjh-NAj-0wbjpAcbDfY9OPSbqHkuqCjzXFQJffMPeuc3waX1EiULUTGRspmRxY4y6e9jwHb_GEb1Nl29TVfv000XXv-5wh7_FWcC1A74bHt8-I-dXl6cnfw2_wkyL7Ux</recordid><startdate>20201109</startdate><enddate>20201109</enddate><creator>Jonker, Hendrik R. A.</creator><creator>Saxena, Krishna</creator><creator>Shcherbakova, Aleksandra</creator><creator>Tiemann, Birgit</creator><creator>Bakker, Hans</creator><creator>Schwalbe, Harald</creator><general>Wiley Subscription Services, Inc</general><general>John Wiley and Sons Inc</general><scope>24P</scope><scope>WIN</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>K9.</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0003-4175-547X</orcidid><orcidid>https://orcid.org/0000-0001-5693-7909</orcidid><orcidid>https://orcid.org/0000-0002-2237-5378</orcidid><orcidid>https://orcid.org/0000-0002-5582-3931</orcidid><orcidid>https://orcid.org/0000-0002-1364-9154</orcidid></search><sort><creationdate>20201109</creationdate><title>NMR Spectroscopic Characterization of the C‐Mannose Conformation in a Thrombospondin Repeat Using a Selective Labeling Approach</title><author>Jonker, Hendrik R. A. ; Saxena, Krishna ; Shcherbakova, Aleksandra ; Tiemann, Birgit ; Bakker, Hans ; Schwalbe, Harald</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5719-67f3a9b36221b8c4a35c8e15eda1ae036e5dfd71c9beb51df58fb2e0d5a5911d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>C-mannosylation</topic><topic>conformation analysis</topic><topic>Glycan</topic><topic>Glycoproteins</topic><topic>Mannose</topic><topic>NMR</topic><topic>NMR spectroscopy</topic><topic>Nuclear magnetic resonance</topic><topic>Polysaccharides</topic><topic>Protein folding</topic><topic>Protein structure</topic><topic>Proteins</topic><topic>selective isotopic labeling</topic><topic>Thrombospondin</topic><topic>Tryptophan</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Jonker, Hendrik R. A.</creatorcontrib><creatorcontrib>Saxena, Krishna</creatorcontrib><creatorcontrib>Shcherbakova, Aleksandra</creatorcontrib><creatorcontrib>Tiemann, Birgit</creatorcontrib><creatorcontrib>Bakker, Hans</creatorcontrib><creatorcontrib>Schwalbe, Harald</creatorcontrib><collection>Wiley Online Library (Open Access Collection)</collection><collection>Wiley Online Library (Open Access Collection)</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Angewandte Chemie International Edition</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Jonker, Hendrik R. A.</au><au>Saxena, Krishna</au><au>Shcherbakova, Aleksandra</au><au>Tiemann, Birgit</au><au>Bakker, Hans</au><au>Schwalbe, Harald</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>NMR Spectroscopic Characterization of the C‐Mannose Conformation in a Thrombospondin Repeat Using a Selective Labeling Approach</atitle><jtitle>Angewandte Chemie International Edition</jtitle><addtitle>Angew Chem Int Ed Engl</addtitle><date>2020-11-09</date><risdate>2020</risdate><volume>59</volume><issue>46</issue><spage>20659</spage><epage>20665</epage><pages>20659-20665</pages><issn>1433-7851</issn><eissn>1521-3773</eissn><abstract>Despite the great interest in glycoproteins, structural information reporting on conformation and dynamics of the sugar moieties are limited. We present a new biochemical method to express proteins with glycans that are selectively labeled with NMR‐active nuclei. We report on the incorporation of 13C‐labeled mannose in the C‐mannosylated UNC‐5 thrombospondin repeat. The conformational landscape of the C‐mannose sugar puckers attached to tryptophan residues of UNC‐5 is characterized by interconversion between the canonical 1C4 state and the B03 / 1S3 state. This flexibility may be essential for protein folding and stabilization. We foresee that this versatile tool to produce proteins with selectively labeled C‐mannose can be applied and adjusted to other systems and modifications and potentially paves a way to advance glycoprotein research by unravelling the dynamical and conformational properties of glycan structures and their interactions.
NMR spectroscopy was used to characterize the C‐mannose conformation in a thrombospondin repeat using a selective labeling approach. The conformational landscape of the C‐mannose sugar puckers attached to tryptophan residues of the netrin receptor UNC‐5 allows interconversion between the canonical 1C4 state and the B03 / 1S3 state.</abstract><cop>Germany</cop><pub>Wiley Subscription Services, Inc</pub><pmid>32745319</pmid><doi>10.1002/anie.202009489</doi><tpages>7</tpages><edition>International ed. in English</edition><orcidid>https://orcid.org/0000-0003-4175-547X</orcidid><orcidid>https://orcid.org/0000-0001-5693-7909</orcidid><orcidid>https://orcid.org/0000-0002-2237-5378</orcidid><orcidid>https://orcid.org/0000-0002-5582-3931</orcidid><orcidid>https://orcid.org/0000-0002-1364-9154</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | C-mannosylation conformation analysis Glycan Glycoproteins Mannose NMR NMR spectroscopy Nuclear magnetic resonance Polysaccharides Protein folding Protein structure Proteins selective isotopic labeling Thrombospondin Tryptophan |
| title | NMR Spectroscopic Characterization of the C‐Mannose Conformation in a Thrombospondin Repeat Using a Selective Labeling Approach |
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