Site‐specific analysis of von Willebrand factor O‐glycosylation
Essentials O‐glycosylation in von Willebrand factor (VWF) is important for its proper function. We report the first detailed site‐specific analysis of the O‐glycosylation in plasma‐derived VWF. All 10 O‐glycosylation sites are occupied, with the disialyl core 1 O‐glycan as the major structure. Core...
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| Veröffentlicht in: | Journal of thrombosis and haemostasis 2016-04, Vol.14 (4), p.733-746 |
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| creator | Solecka, B. A. Weise, C. Laffan, M. A. Kannicht, C. |
| description | Essentials
O‐glycosylation in von Willebrand factor (VWF) is important for its proper function.
We report the first detailed site‐specific analysis of the O‐glycosylation in plasma‐derived VWF.
All 10 O‐glycosylation sites are occupied, with the disialyl core 1 O‐glycan as the major structure.
Core 2 O‐glycan is unevenly distributed and, sulfated O‐glycans and Tn antigen have been found.
Summary
Background
O‐glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O‐glycosylation sites.
Objectives
The aim of this study was the site‐specific analysis of VWF O‐glycosylation.
Methods
Tryptic VWF‐O‐glycopeptides were isolated by lectin affinity chromatography and/or by reverse‐phase high‐performance liquid chromatography. Subsequently, the purified glycopeptides were analyzed by glycosidase digestion and mass spectrometry.
Results
We found that all 10 predicted O‐glycosylation sites in VWF are occupied. The majority of the glycan structures on all glycosylation sites is represented by disialyl core 1 O‐glycan. The presence of core 2 O‐glycan was also confirmed; interestingly, this structure was not evenly distributed among all 10 glycosylation sites. Analysis of the glycopeptides flanking the A1 domain revealed that generally more core‐2‐type O‐glycan was present on the C‐terminal Cluster 2 glycopeptide (encompassing T1468, T1477, S1486 and T1487) compared with the N‐terminal Cluster 1 glycopeptide (encompassing T1248, T1255, T1256 and S1263). Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T2298 in the C1 domain. In addition, we identify sulfation of core 2 O‐glycans and the presence of the rare Tn antigen.
Conclusions
This is the first study to describe the qualitative and semi‐quantitative distribution of O‐glycan structures on all 10 O‐glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O‐glycosylation in VWF. |
| doi_str_mv | 10.1111/jth.13260 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_1780510259</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1780510259</sourcerecordid><originalsourceid>FETCH-LOGICAL-c4540-6ae0875d725fd435de93b53550864b483ef135317c2d57bc64643d2e9b369feb3</originalsourceid><addsrcrecordid>eNp10L1OwzAUBWALgWgpDLwAisQCQ1o7_kkyogooqBIDRYyW4zjgyo2LnYCy8Qg8I0-CIS0DEne5d_h0pHsAOEZwjMJMls3zGOGEwR0wRBRncZphtru9c4wH4MD7JYQopwncB4OEpRmhmAzB9F436vP9w6-V1JWWkaiF6bz2ka2iV1tHj9oYVThRl1ElZGNddBf4k-mk9Z0Rjbb1IdirhPHqaLNH4OHqcjGdxfO765vpxTyWhBIYM6FgltIyTWhVEkxLleOCYkphxkhBMqwqhClGqUxKmhaSEUZwmai8wCyvVIFH4KzPXTv70irf8JX2UhkjamVbz1GaQYpgQvNAT__QpW1deK1XEDJKcVDnvZLOeu9UxddOr4TrOIL8u1kemuU_zQZ7sklsi5Uqf-W2ygAmPXjTRnX_J_HbxayP_AIsgoKF</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1780006553</pqid></control><display><type>article</type><title>Site‐specific analysis of von Willebrand factor O‐glycosylation</title><source>MEDLINE</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>Alma/SFX Local Collection</source><creator>Solecka, B. A. ; Weise, C. ; Laffan, M. A. ; Kannicht, C.</creator><creatorcontrib>Solecka, B. A. ; Weise, C. ; Laffan, M. A. ; Kannicht, C.</creatorcontrib><description>Essentials
O‐glycosylation in von Willebrand factor (VWF) is important for its proper function.
We report the first detailed site‐specific analysis of the O‐glycosylation in plasma‐derived VWF.
All 10 O‐glycosylation sites are occupied, with the disialyl core 1 O‐glycan as the major structure.
Core 2 O‐glycan is unevenly distributed and, sulfated O‐glycans and Tn antigen have been found.
Summary
Background
O‐glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O‐glycosylation sites.
Objectives
The aim of this study was the site‐specific analysis of VWF O‐glycosylation.
Methods
Tryptic VWF‐O‐glycopeptides were isolated by lectin affinity chromatography and/or by reverse‐phase high‐performance liquid chromatography. Subsequently, the purified glycopeptides were analyzed by glycosidase digestion and mass spectrometry.
Results
We found that all 10 predicted O‐glycosylation sites in VWF are occupied. The majority of the glycan structures on all glycosylation sites is represented by disialyl core 1 O‐glycan. The presence of core 2 O‐glycan was also confirmed; interestingly, this structure was not evenly distributed among all 10 glycosylation sites. Analysis of the glycopeptides flanking the A1 domain revealed that generally more core‐2‐type O‐glycan was present on the C‐terminal Cluster 2 glycopeptide (encompassing T1468, T1477, S1486 and T1487) compared with the N‐terminal Cluster 1 glycopeptide (encompassing T1248, T1255, T1256 and S1263). Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T2298 in the C1 domain. In addition, we identify sulfation of core 2 O‐glycans and the presence of the rare Tn antigen.
Conclusions
This is the first study to describe the qualitative and semi‐quantitative distribution of O‐glycan structures on all 10 O‐glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O‐glycosylation in VWF.</description><identifier>ISSN: 1538-7933</identifier><identifier>ISSN: 1538-7836</identifier><identifier>EISSN: 1538-7836</identifier><identifier>DOI: 10.1111/jth.13260</identifier><identifier>PMID: 26784534</identifier><language>eng</language><publisher>England: Elsevier Limited</publisher><subject>Antigens, Tumor-Associated, Carbohydrate - chemistry ; Chromatography, Affinity ; Chromatography, High Pressure Liquid ; glycopeptides ; Glycopeptides - chemistry ; Glycoside Hydrolases - chemistry ; Glycosylation ; Humans ; Jacalin ; Lectins - chemistry ; Mass Spectrometry ; matrix‐assisted laser desorption‐ionization mass spectrometry ; Plant Lectins - chemistry ; Polysaccharides - chemistry ; Protein Domains ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; von Willebrand factor ; von Willebrand Factor - chemistry</subject><ispartof>Journal of thrombosis and haemostasis, 2016-04, Vol.14 (4), p.733-746</ispartof><rights>2016 International Society on Thrombosis and Haemostasis</rights><rights>2016 International Society on Thrombosis and Haemostasis.</rights><rights>Copyright © 2016 International Society on Thrombosis and Haemostasis</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4540-6ae0875d725fd435de93b53550864b483ef135317c2d57bc64643d2e9b369feb3</citedby><cites>FETCH-LOGICAL-c4540-6ae0875d725fd435de93b53550864b483ef135317c2d57bc64643d2e9b369feb3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26784534$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Solecka, B. A.</creatorcontrib><creatorcontrib>Weise, C.</creatorcontrib><creatorcontrib>Laffan, M. A.</creatorcontrib><creatorcontrib>Kannicht, C.</creatorcontrib><title>Site‐specific analysis of von Willebrand factor O‐glycosylation</title><title>Journal of thrombosis and haemostasis</title><addtitle>J Thromb Haemost</addtitle><description>Essentials
O‐glycosylation in von Willebrand factor (VWF) is important for its proper function.
We report the first detailed site‐specific analysis of the O‐glycosylation in plasma‐derived VWF.
All 10 O‐glycosylation sites are occupied, with the disialyl core 1 O‐glycan as the major structure.
Core 2 O‐glycan is unevenly distributed and, sulfated O‐glycans and Tn antigen have been found.
Summary
Background
O‐glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O‐glycosylation sites.
Objectives
The aim of this study was the site‐specific analysis of VWF O‐glycosylation.
Methods
Tryptic VWF‐O‐glycopeptides were isolated by lectin affinity chromatography and/or by reverse‐phase high‐performance liquid chromatography. Subsequently, the purified glycopeptides were analyzed by glycosidase digestion and mass spectrometry.
Results
We found that all 10 predicted O‐glycosylation sites in VWF are occupied. The majority of the glycan structures on all glycosylation sites is represented by disialyl core 1 O‐glycan. The presence of core 2 O‐glycan was also confirmed; interestingly, this structure was not evenly distributed among all 10 glycosylation sites. Analysis of the glycopeptides flanking the A1 domain revealed that generally more core‐2‐type O‐glycan was present on the C‐terminal Cluster 2 glycopeptide (encompassing T1468, T1477, S1486 and T1487) compared with the N‐terminal Cluster 1 glycopeptide (encompassing T1248, T1255, T1256 and S1263). Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T2298 in the C1 domain. In addition, we identify sulfation of core 2 O‐glycans and the presence of the rare Tn antigen.
Conclusions
This is the first study to describe the qualitative and semi‐quantitative distribution of O‐glycan structures on all 10 O‐glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O‐glycosylation in VWF.</description><subject>Antigens, Tumor-Associated, Carbohydrate - chemistry</subject><subject>Chromatography, Affinity</subject><subject>Chromatography, High Pressure Liquid</subject><subject>glycopeptides</subject><subject>Glycopeptides - chemistry</subject><subject>Glycoside Hydrolases - chemistry</subject><subject>Glycosylation</subject><subject>Humans</subject><subject>Jacalin</subject><subject>Lectins - chemistry</subject><subject>Mass Spectrometry</subject><subject>matrix‐assisted laser desorption‐ionization mass spectrometry</subject><subject>Plant Lectins - chemistry</subject><subject>Polysaccharides - chemistry</subject><subject>Protein Domains</subject><subject>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</subject><subject>von Willebrand factor</subject><subject>von Willebrand Factor - chemistry</subject><issn>1538-7933</issn><issn>1538-7836</issn><issn>1538-7836</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp10L1OwzAUBWALgWgpDLwAisQCQ1o7_kkyogooqBIDRYyW4zjgyo2LnYCy8Qg8I0-CIS0DEne5d_h0pHsAOEZwjMJMls3zGOGEwR0wRBRncZphtru9c4wH4MD7JYQopwncB4OEpRmhmAzB9F436vP9w6-V1JWWkaiF6bz2ka2iV1tHj9oYVThRl1ElZGNddBf4k-mk9Z0Rjbb1IdirhPHqaLNH4OHqcjGdxfO765vpxTyWhBIYM6FgltIyTWhVEkxLleOCYkphxkhBMqwqhClGqUxKmhaSEUZwmai8wCyvVIFH4KzPXTv70irf8JX2UhkjamVbz1GaQYpgQvNAT__QpW1deK1XEDJKcVDnvZLOeu9UxddOr4TrOIL8u1kemuU_zQZ7sklsi5Uqf-W2ygAmPXjTRnX_J_HbxayP_AIsgoKF</recordid><startdate>201604</startdate><enddate>201604</enddate><creator>Solecka, B. A.</creator><creator>Weise, C.</creator><creator>Laffan, M. A.</creator><creator>Kannicht, C.</creator><general>Elsevier Limited</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>7T5</scope><scope>H94</scope><scope>K9.</scope><scope>7X8</scope></search><sort><creationdate>201604</creationdate><title>Site‐specific analysis of von Willebrand factor O‐glycosylation</title><author>Solecka, B. A. ; Weise, C. ; Laffan, M. A. ; Kannicht, C.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4540-6ae0875d725fd435de93b53550864b483ef135317c2d57bc64643d2e9b369feb3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Antigens, Tumor-Associated, Carbohydrate - chemistry</topic><topic>Chromatography, Affinity</topic><topic>Chromatography, High Pressure Liquid</topic><topic>glycopeptides</topic><topic>Glycopeptides - chemistry</topic><topic>Glycoside Hydrolases - chemistry</topic><topic>Glycosylation</topic><topic>Humans</topic><topic>Jacalin</topic><topic>Lectins - chemistry</topic><topic>Mass Spectrometry</topic><topic>matrix‐assisted laser desorption‐ionization mass spectrometry</topic><topic>Plant Lectins - chemistry</topic><topic>Polysaccharides - chemistry</topic><topic>Protein Domains</topic><topic>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</topic><topic>von Willebrand factor</topic><topic>von Willebrand Factor - chemistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Solecka, B. A.</creatorcontrib><creatorcontrib>Weise, C.</creatorcontrib><creatorcontrib>Laffan, M. A.</creatorcontrib><creatorcontrib>Kannicht, C.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of thrombosis and haemostasis</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Solecka, B. A.</au><au>Weise, C.</au><au>Laffan, M. A.</au><au>Kannicht, C.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Site‐specific analysis of von Willebrand factor O‐glycosylation</atitle><jtitle>Journal of thrombosis and haemostasis</jtitle><addtitle>J Thromb Haemost</addtitle><date>2016-04</date><risdate>2016</risdate><volume>14</volume><issue>4</issue><spage>733</spage><epage>746</epage><pages>733-746</pages><issn>1538-7933</issn><issn>1538-7836</issn><eissn>1538-7836</eissn><abstract>Essentials
O‐glycosylation in von Willebrand factor (VWF) is important for its proper function.
We report the first detailed site‐specific analysis of the O‐glycosylation in plasma‐derived VWF.
All 10 O‐glycosylation sites are occupied, with the disialyl core 1 O‐glycan as the major structure.
Core 2 O‐glycan is unevenly distributed and, sulfated O‐glycans and Tn antigen have been found.
Summary
Background
O‐glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O‐glycosylation sites.
Objectives
The aim of this study was the site‐specific analysis of VWF O‐glycosylation.
Methods
Tryptic VWF‐O‐glycopeptides were isolated by lectin affinity chromatography and/or by reverse‐phase high‐performance liquid chromatography. Subsequently, the purified glycopeptides were analyzed by glycosidase digestion and mass spectrometry.
Results
We found that all 10 predicted O‐glycosylation sites in VWF are occupied. The majority of the glycan structures on all glycosylation sites is represented by disialyl core 1 O‐glycan. The presence of core 2 O‐glycan was also confirmed; interestingly, this structure was not evenly distributed among all 10 glycosylation sites. Analysis of the glycopeptides flanking the A1 domain revealed that generally more core‐2‐type O‐glycan was present on the C‐terminal Cluster 2 glycopeptide (encompassing T1468, T1477, S1486 and T1487) compared with the N‐terminal Cluster 1 glycopeptide (encompassing T1248, T1255, T1256 and S1263). Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T2298 in the C1 domain. In addition, we identify sulfation of core 2 O‐glycans and the presence of the rare Tn antigen.
Conclusions
This is the first study to describe the qualitative and semi‐quantitative distribution of O‐glycan structures on all 10 O‐glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O‐glycosylation in VWF.</abstract><cop>England</cop><pub>Elsevier Limited</pub><pmid>26784534</pmid><doi>10.1111/jth.13260</doi><tpages>14</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Journal of thrombosis and haemostasis, 2016-04, Vol.14 (4), p.733-746 |
| issn | 1538-7933 1538-7836 1538-7836 |
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| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Antigens, Tumor-Associated, Carbohydrate - chemistry Chromatography, Affinity Chromatography, High Pressure Liquid glycopeptides Glycopeptides - chemistry Glycoside Hydrolases - chemistry Glycosylation Humans Jacalin Lectins - chemistry Mass Spectrometry matrix‐assisted laser desorption‐ionization mass spectrometry Plant Lectins - chemistry Polysaccharides - chemistry Protein Domains Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization von Willebrand factor von Willebrand Factor - chemistry |
| title | Site‐specific analysis of von Willebrand factor O‐glycosylation |
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