Mass Spectrometry-Based Shotgun Glycomics for Discovery of Natural Ligands of Glycan-Binding Proteins
Glycans attached to lipids and membrane-bound and secreted proteins and peptides mediate many important physiological and pathophysiological processes through interactions with glycan-binding proteins (GBPs). However, uncovering functional glycan ligands is challenging due to the large number of nat...
Gespeichert in:
| Veröffentlicht in: | Analytical chemistry (Washington) 2020-10, Vol.92 (20), p.14012-14020 |
|---|---|
| Hauptverfasser: | , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 14020 |
|---|---|
| container_issue | 20 |
| container_start_page | 14012 |
| container_title | Analytical chemistry (Washington) |
| container_volume | 92 |
| creator | Park, Heajin Jung, Jaesoo Rodrigues, Emily Kitova, Elena N Macauley, Matthew S Klassen, John S |
| description | Glycans attached to lipids and membrane-bound and secreted proteins and peptides mediate many important physiological and pathophysiological processes through interactions with glycan-binding proteins (GBPs). However, uncovering functional glycan ligands is challenging due to the large number of naturally occurring glycan structures, the limited availability of glycans in their purified form, the low affinities of GBP-glycan interactions, and limitations in existing binding assays. This work explores the application of catch-and-release electrospray ionization mass spectrometry (CaR-ESI-MS) for screening libraries of
-glycans derived from natural sources. The assay was tested by screening a small-defined library of complex
-glycans at equimolar concentrations against plant and human GBPs with known specificities for either α2-3- or α2-6-linked sialosides, with affinities in the millimolar to micromolar range. Validation experiments, performed in negative ion mode, revealed that bound
-glycan ligands are readily released, as intact deprotonated ions, from GBPs in the gas phase using collision-induced dissociation. Moreover, the relative abundances of the released ligands closely match their solution affinities. The results obtained for a natural
-glycan library produced from cultured immune cells serve to highlight the ease with which CaR-ESI-MS can screen complex mixtures of
-glycans for interactions. Additionally, scaling the relative abundances of released glycan ligands according to their relative abundances in solution, as determined by hydrophilic interaction-ultrahigh-performance liquid chromatography of the fluorescently labeled library, allows the relative affinities of glycan ligands to be ranked. |
| doi_str_mv | 10.1021/acs.analchem.0c02931 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_2443881959</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2443881959</sourcerecordid><originalsourceid>FETCH-LOGICAL-c335t-65075cda88d4e2d93cd0d06731b0ad9db747037416a7595ea5b7b08e5137d1ba3</originalsourceid><addsrcrecordid>eNpdkU1Lw0AQhhdRbP34ByILXrykzmaz2eRoq1ahfkD1HCa72zYlydbdRMi_N6WtB08Dw_O-DPMQcsVgxCBkd6j8CGss1cpUI1AQppwdkSETIQRxkoTHZAgAPAglwICceb8GYAxYfEoGvIfjGOIhMa_oPZ1vjGqcrUzjumCM3mg6X9lm2dZ0WnbKVoXydGEdfSi8sj_GddQu6Bs2rcOSzool1tpvV1sa62Bc1Lqol_TD2cYUtb8gJwssvbncz3Py9fT4OXkOZu_Tl8n9LFCciyaIBUihNCaJjkyoU640aIglZzmgTnUuIwlcRixGKVJhUOQyh8QIxqVmOfJzcrvr3Tj73RrfZFV_sClLrI1tfRZGEU8Sloq0R2_-oWvbuv6fW0pEPcPSsKeiHaWc9d6ZRbZxRYWuyxhkWw1ZryE7aMj2GvrY9b68zSuj_0KHv_Nf3FmGjg</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2454195192</pqid></control><display><type>article</type><title>Mass Spectrometry-Based Shotgun Glycomics for Discovery of Natural Ligands of Glycan-Binding Proteins</title><source>ACS Publications</source><source>MEDLINE</source><creator>Park, Heajin ; Jung, Jaesoo ; Rodrigues, Emily ; Kitova, Elena N ; Macauley, Matthew S ; Klassen, John S</creator><creatorcontrib>Park, Heajin ; Jung, Jaesoo ; Rodrigues, Emily ; Kitova, Elena N ; Macauley, Matthew S ; Klassen, John S</creatorcontrib><description>Glycans attached to lipids and membrane-bound and secreted proteins and peptides mediate many important physiological and pathophysiological processes through interactions with glycan-binding proteins (GBPs). However, uncovering functional glycan ligands is challenging due to the large number of naturally occurring glycan structures, the limited availability of glycans in their purified form, the low affinities of GBP-glycan interactions, and limitations in existing binding assays. This work explores the application of catch-and-release electrospray ionization mass spectrometry (CaR-ESI-MS) for screening libraries of
-glycans derived from natural sources. The assay was tested by screening a small-defined library of complex
-glycans at equimolar concentrations against plant and human GBPs with known specificities for either α2-3- or α2-6-linked sialosides, with affinities in the millimolar to micromolar range. Validation experiments, performed in negative ion mode, revealed that bound
-glycan ligands are readily released, as intact deprotonated ions, from GBPs in the gas phase using collision-induced dissociation. Moreover, the relative abundances of the released ligands closely match their solution affinities. The results obtained for a natural
-glycan library produced from cultured immune cells serve to highlight the ease with which CaR-ESI-MS can screen complex mixtures of
-glycans for interactions. Additionally, scaling the relative abundances of released glycan ligands according to their relative abundances in solution, as determined by hydrophilic interaction-ultrahigh-performance liquid chromatography of the fluorescently labeled library, allows the relative affinities of glycan ligands to be ranked.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/acs.analchem.0c02931</identifier><identifier>PMID: 32936606</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Abundance ; Affinity ; Agglutinins - chemistry ; Agglutinins - metabolism ; Binding ; Chemistry ; Chromatography, High Pressure Liquid ; Glycan ; Glycomics - methods ; Humans ; Immune system ; Ionization ; Ions ; Libraries ; Ligands ; Lipids ; Liquid chromatography ; Mass spectrometry ; Mass spectroscopy ; N-glycans ; Negative ions ; Peptides ; Plant Lectins - chemistry ; Plant Lectins - metabolism ; Polysaccharides ; Polysaccharides - analysis ; Polysaccharides - metabolism ; Protein Binding ; Proteins ; Sambucus nigra - metabolism ; Scientific imaging ; Screening ; Shotguns ; Sialic Acid Binding Ig-like Lectin 2 - chemistry ; Sialic Acid Binding Ig-like Lectin 2 - metabolism ; Spectrometry, Mass, Electrospray Ionization ; Spectroscopy ; Vapor phases</subject><ispartof>Analytical chemistry (Washington), 2020-10, Vol.92 (20), p.14012-14020</ispartof><rights>Copyright American Chemical Society Oct 20, 2020</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c335t-65075cda88d4e2d93cd0d06731b0ad9db747037416a7595ea5b7b08e5137d1ba3</citedby><cites>FETCH-LOGICAL-c335t-65075cda88d4e2d93cd0d06731b0ad9db747037416a7595ea5b7b08e5137d1ba3</cites><orcidid>0000-0003-4579-1048 ; 0000-0002-3389-7112</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,2752,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32936606$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Park, Heajin</creatorcontrib><creatorcontrib>Jung, Jaesoo</creatorcontrib><creatorcontrib>Rodrigues, Emily</creatorcontrib><creatorcontrib>Kitova, Elena N</creatorcontrib><creatorcontrib>Macauley, Matthew S</creatorcontrib><creatorcontrib>Klassen, John S</creatorcontrib><title>Mass Spectrometry-Based Shotgun Glycomics for Discovery of Natural Ligands of Glycan-Binding Proteins</title><title>Analytical chemistry (Washington)</title><addtitle>Anal Chem</addtitle><description>Glycans attached to lipids and membrane-bound and secreted proteins and peptides mediate many important physiological and pathophysiological processes through interactions with glycan-binding proteins (GBPs). However, uncovering functional glycan ligands is challenging due to the large number of naturally occurring glycan structures, the limited availability of glycans in their purified form, the low affinities of GBP-glycan interactions, and limitations in existing binding assays. This work explores the application of catch-and-release electrospray ionization mass spectrometry (CaR-ESI-MS) for screening libraries of
-glycans derived from natural sources. The assay was tested by screening a small-defined library of complex
-glycans at equimolar concentrations against plant and human GBPs with known specificities for either α2-3- or α2-6-linked sialosides, with affinities in the millimolar to micromolar range. Validation experiments, performed in negative ion mode, revealed that bound
-glycan ligands are readily released, as intact deprotonated ions, from GBPs in the gas phase using collision-induced dissociation. Moreover, the relative abundances of the released ligands closely match their solution affinities. The results obtained for a natural
-glycan library produced from cultured immune cells serve to highlight the ease with which CaR-ESI-MS can screen complex mixtures of
-glycans for interactions. Additionally, scaling the relative abundances of released glycan ligands according to their relative abundances in solution, as determined by hydrophilic interaction-ultrahigh-performance liquid chromatography of the fluorescently labeled library, allows the relative affinities of glycan ligands to be ranked.</description><subject>Abundance</subject><subject>Affinity</subject><subject>Agglutinins - chemistry</subject><subject>Agglutinins - metabolism</subject><subject>Binding</subject><subject>Chemistry</subject><subject>Chromatography, High Pressure Liquid</subject><subject>Glycan</subject><subject>Glycomics - methods</subject><subject>Humans</subject><subject>Immune system</subject><subject>Ionization</subject><subject>Ions</subject><subject>Libraries</subject><subject>Ligands</subject><subject>Lipids</subject><subject>Liquid chromatography</subject><subject>Mass spectrometry</subject><subject>Mass spectroscopy</subject><subject>N-glycans</subject><subject>Negative ions</subject><subject>Peptides</subject><subject>Plant Lectins - chemistry</subject><subject>Plant Lectins - metabolism</subject><subject>Polysaccharides</subject><subject>Polysaccharides - analysis</subject><subject>Polysaccharides - metabolism</subject><subject>Protein Binding</subject><subject>Proteins</subject><subject>Sambucus nigra - metabolism</subject><subject>Scientific imaging</subject><subject>Screening</subject><subject>Shotguns</subject><subject>Sialic Acid Binding Ig-like Lectin 2 - chemistry</subject><subject>Sialic Acid Binding Ig-like Lectin 2 - metabolism</subject><subject>Spectrometry, Mass, Electrospray Ionization</subject><subject>Spectroscopy</subject><subject>Vapor phases</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdkU1Lw0AQhhdRbP34ByILXrykzmaz2eRoq1ahfkD1HCa72zYlydbdRMi_N6WtB08Dw_O-DPMQcsVgxCBkd6j8CGss1cpUI1AQppwdkSETIQRxkoTHZAgAPAglwICceb8GYAxYfEoGvIfjGOIhMa_oPZ1vjGqcrUzjumCM3mg6X9lm2dZ0WnbKVoXydGEdfSi8sj_GddQu6Bs2rcOSzool1tpvV1sa62Bc1Lqol_TD2cYUtb8gJwssvbncz3Py9fT4OXkOZu_Tl8n9LFCciyaIBUihNCaJjkyoU640aIglZzmgTnUuIwlcRixGKVJhUOQyh8QIxqVmOfJzcrvr3Tj73RrfZFV_sClLrI1tfRZGEU8Sloq0R2_-oWvbuv6fW0pEPcPSsKeiHaWc9d6ZRbZxRYWuyxhkWw1ZryE7aMj2GvrY9b68zSuj_0KHv_Nf3FmGjg</recordid><startdate>20201020</startdate><enddate>20201020</enddate><creator>Park, Heajin</creator><creator>Jung, Jaesoo</creator><creator>Rodrigues, Emily</creator><creator>Kitova, Elena N</creator><creator>Macauley, Matthew S</creator><creator>Klassen, John S</creator><general>American Chemical Society</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>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0003-4579-1048</orcidid><orcidid>https://orcid.org/0000-0002-3389-7112</orcidid></search><sort><creationdate>20201020</creationdate><title>Mass Spectrometry-Based Shotgun Glycomics for Discovery of Natural Ligands of Glycan-Binding Proteins</title><author>Park, Heajin ; Jung, Jaesoo ; Rodrigues, Emily ; Kitova, Elena N ; Macauley, Matthew S ; Klassen, John S</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c335t-65075cda88d4e2d93cd0d06731b0ad9db747037416a7595ea5b7b08e5137d1ba3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Abundance</topic><topic>Affinity</topic><topic>Agglutinins - chemistry</topic><topic>Agglutinins - metabolism</topic><topic>Binding</topic><topic>Chemistry</topic><topic>Chromatography, High Pressure Liquid</topic><topic>Glycan</topic><topic>Glycomics - methods</topic><topic>Humans</topic><topic>Immune system</topic><topic>Ionization</topic><topic>Ions</topic><topic>Libraries</topic><topic>Ligands</topic><topic>Lipids</topic><topic>Liquid chromatography</topic><topic>Mass spectrometry</topic><topic>Mass spectroscopy</topic><topic>N-glycans</topic><topic>Negative ions</topic><topic>Peptides</topic><topic>Plant Lectins - chemistry</topic><topic>Plant Lectins - metabolism</topic><topic>Polysaccharides</topic><topic>Polysaccharides - analysis</topic><topic>Polysaccharides - metabolism</topic><topic>Protein Binding</topic><topic>Proteins</topic><topic>Sambucus nigra - metabolism</topic><topic>Scientific imaging</topic><topic>Screening</topic><topic>Shotguns</topic><topic>Sialic Acid Binding Ig-like Lectin 2 - chemistry</topic><topic>Sialic Acid Binding Ig-like Lectin 2 - metabolism</topic><topic>Spectrometry, Mass, Electrospray Ionization</topic><topic>Spectroscopy</topic><topic>Vapor phases</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Park, Heajin</creatorcontrib><creatorcontrib>Jung, Jaesoo</creatorcontrib><creatorcontrib>Rodrigues, Emily</creatorcontrib><creatorcontrib>Kitova, Elena N</creatorcontrib><creatorcontrib>Macauley, Matthew S</creatorcontrib><creatorcontrib>Klassen, John S</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Park, Heajin</au><au>Jung, Jaesoo</au><au>Rodrigues, Emily</au><au>Kitova, Elena N</au><au>Macauley, Matthew S</au><au>Klassen, John S</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Mass Spectrometry-Based Shotgun Glycomics for Discovery of Natural Ligands of Glycan-Binding Proteins</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal Chem</addtitle><date>2020-10-20</date><risdate>2020</risdate><volume>92</volume><issue>20</issue><spage>14012</spage><epage>14020</epage><pages>14012-14020</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><abstract>Glycans attached to lipids and membrane-bound and secreted proteins and peptides mediate many important physiological and pathophysiological processes through interactions with glycan-binding proteins (GBPs). However, uncovering functional glycan ligands is challenging due to the large number of naturally occurring glycan structures, the limited availability of glycans in their purified form, the low affinities of GBP-glycan interactions, and limitations in existing binding assays. This work explores the application of catch-and-release electrospray ionization mass spectrometry (CaR-ESI-MS) for screening libraries of
-glycans derived from natural sources. The assay was tested by screening a small-defined library of complex
-glycans at equimolar concentrations against plant and human GBPs with known specificities for either α2-3- or α2-6-linked sialosides, with affinities in the millimolar to micromolar range. Validation experiments, performed in negative ion mode, revealed that bound
-glycan ligands are readily released, as intact deprotonated ions, from GBPs in the gas phase using collision-induced dissociation. Moreover, the relative abundances of the released ligands closely match their solution affinities. The results obtained for a natural
-glycan library produced from cultured immune cells serve to highlight the ease with which CaR-ESI-MS can screen complex mixtures of
-glycans for interactions. Additionally, scaling the relative abundances of released glycan ligands according to their relative abundances in solution, as determined by hydrophilic interaction-ultrahigh-performance liquid chromatography of the fluorescently labeled library, allows the relative affinities of glycan ligands to be ranked.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>32936606</pmid><doi>10.1021/acs.analchem.0c02931</doi><tpages>9</tpages><orcidid>https://orcid.org/0000-0003-4579-1048</orcidid><orcidid>https://orcid.org/0000-0002-3389-7112</orcidid></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0003-2700 |
| ispartof | Analytical chemistry (Washington), 2020-10, Vol.92 (20), p.14012-14020 |
| issn | 0003-2700 1520-6882 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_2443881959 |
| source | ACS Publications; MEDLINE |
| subjects | Abundance Affinity Agglutinins - chemistry Agglutinins - metabolism Binding Chemistry Chromatography, High Pressure Liquid Glycan Glycomics - methods Humans Immune system Ionization Ions Libraries Ligands Lipids Liquid chromatography Mass spectrometry Mass spectroscopy N-glycans Negative ions Peptides Plant Lectins - chemistry Plant Lectins - metabolism Polysaccharides Polysaccharides - analysis Polysaccharides - metabolism Protein Binding Proteins Sambucus nigra - metabolism Scientific imaging Screening Shotguns Sialic Acid Binding Ig-like Lectin 2 - chemistry Sialic Acid Binding Ig-like Lectin 2 - metabolism Spectrometry, Mass, Electrospray Ionization Spectroscopy Vapor phases |
| title | Mass Spectrometry-Based Shotgun Glycomics for Discovery of Natural Ligands of Glycan-Binding Proteins |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-21T20%3A16%3A19IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Mass%20Spectrometry-Based%20Shotgun%20Glycomics%20for%20Discovery%20of%20Natural%20Ligands%20of%20Glycan-Binding%20Proteins&rft.jtitle=Analytical%20chemistry%20(Washington)&rft.au=Park,%20Heajin&rft.date=2020-10-20&rft.volume=92&rft.issue=20&rft.spage=14012&rft.epage=14020&rft.pages=14012-14020&rft.issn=0003-2700&rft.eissn=1520-6882&rft_id=info:doi/10.1021/acs.analchem.0c02931&rft_dat=%3Cproquest_cross%3E2443881959%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2454195192&rft_id=info:pmid/32936606&rfr_iscdi=true |