The relationship between glycan structures and expression levels of an endoplasmic reticulum-resident glycoprotein, UDP-glucose: Glycoprotein glucosyltransferase 1
In this article, we report a relationship between glycan structures and expression levels of a recombinant ER-resident glycoprotein, uridine 5′-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1). The function of glycan structures attached to a glycoprotein is actively studied; however, th...
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| Veröffentlicht in: | Biochemical and biophysical research communications 2015-06, Vol.462 (1), p.58-63 |
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| container_title | Biochemical and biophysical research communications |
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| creator | Daikoku, Shusaku Seko, Akira Son, Sang-Hyun Suzuki, Katsuhiko Ito, Yukishige Kanie, Osamu |
| description | In this article, we report a relationship between glycan structures and expression levels of a recombinant ER-resident glycoprotein, uridine 5′-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1). The function of glycan structures attached to a glycoprotein is actively studied; however, the glycan structures of recombinant, and not endogenous, glycoproteins have not been examined. In this study, we indicate a relationship between the glycan structure and the level of protein expression. Expression levels were controlled utilizing a series of vectors (pFN21K, pFN22K, pFN23K, and pFN24K HaloTag CMV Flexi Vectors). Qualitative and semi-quantitative confirmation of glycan structures was achieved with tandem mass spectrometry. The results of this study indicate that glycan structures are similar to endogenous glycans at low expression levels.
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•The glycan profiles of glycoproteins were investigated using four expression vectors.•At low expression levels, glycan structures were similar to endogenous glycans.•The N-linked glycan structure of UGGT1 is dependent on the peptide expression level. |
| doi_str_mv | 10.1016/j.bbrc.2015.04.105 |
| format | Article |
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[Display omitted]
•The glycan profiles of glycoproteins were investigated using four expression vectors.•At low expression levels, glycan structures were similar to endogenous glycans.•The N-linked glycan structure of UGGT1 is dependent on the peptide expression level.</description><identifier>ISSN: 0006-291X</identifier><identifier>EISSN: 1090-2104</identifier><identifier>DOI: 10.1016/j.bbrc.2015.04.105</identifier><identifier>PMID: 25935482</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Chromatography, Liquid - methods ; Electrophoresis, Polyacrylamide Gel ; Endoplasmic Reticulum - metabolism ; Expression level ; Glucosyltransferases - genetics ; Glucosyltransferases - metabolism ; Glycoproteins - genetics ; Glycoproteins - metabolism ; HEK293 Cells ; Humans ; Mass Spectrometry - methods ; NanoLC–tandem MS ; Polysaccharides - chemistry ; Polysaccharides - metabolism ; Recombinant Proteins - metabolism ; Uridine 5′-diphosphate-glucose: glycoprotein glucosyltransferase 1</subject><ispartof>Biochemical and biophysical research communications, 2015-06, Vol.462 (1), p.58-63</ispartof><rights>2015 Elsevier Inc.</rights><rights>Copyright © 2015 Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c422t-f6c9798804d8716d5ed9c8fd064c2a4b690d16e39a4a281615259b90ce3e6c093</citedby><cites>FETCH-LOGICAL-c422t-f6c9798804d8716d5ed9c8fd064c2a4b690d16e39a4a281615259b90ce3e6c093</cites><orcidid>0000-0003-4147-6868</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0006291X15008098$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25935482$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Daikoku, Shusaku</creatorcontrib><creatorcontrib>Seko, Akira</creatorcontrib><creatorcontrib>Son, Sang-Hyun</creatorcontrib><creatorcontrib>Suzuki, Katsuhiko</creatorcontrib><creatorcontrib>Ito, Yukishige</creatorcontrib><creatorcontrib>Kanie, Osamu</creatorcontrib><title>The relationship between glycan structures and expression levels of an endoplasmic reticulum-resident glycoprotein, UDP-glucose: Glycoprotein glucosyltransferase 1</title><title>Biochemical and biophysical research communications</title><addtitle>Biochem Biophys Res Commun</addtitle><description>In this article, we report a relationship between glycan structures and expression levels of a recombinant ER-resident glycoprotein, uridine 5′-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1). The function of glycan structures attached to a glycoprotein is actively studied; however, the glycan structures of recombinant, and not endogenous, glycoproteins have not been examined. In this study, we indicate a relationship between the glycan structure and the level of protein expression. Expression levels were controlled utilizing a series of vectors (pFN21K, pFN22K, pFN23K, and pFN24K HaloTag CMV Flexi Vectors). Qualitative and semi-quantitative confirmation of glycan structures was achieved with tandem mass spectrometry. The results of this study indicate that glycan structures are similar to endogenous glycans at low expression levels.
[Display omitted]
•The glycan profiles of glycoproteins were investigated using four expression vectors.•At low expression levels, glycan structures were similar to endogenous glycans.•The N-linked glycan structure of UGGT1 is dependent on the peptide expression level.</description><subject>Chromatography, Liquid - methods</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Endoplasmic Reticulum - metabolism</subject><subject>Expression level</subject><subject>Glucosyltransferases - genetics</subject><subject>Glucosyltransferases - metabolism</subject><subject>Glycoproteins - genetics</subject><subject>Glycoproteins - metabolism</subject><subject>HEK293 Cells</subject><subject>Humans</subject><subject>Mass Spectrometry - methods</subject><subject>NanoLC–tandem MS</subject><subject>Polysaccharides - chemistry</subject><subject>Polysaccharides - metabolism</subject><subject>Recombinant Proteins - metabolism</subject><subject>Uridine 5′-diphosphate-glucose: glycoprotein glucosyltransferase 1</subject><issn>0006-291X</issn><issn>1090-2104</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kb1uFDEURi0EIkvgBSiQSwpmufZ4vGNEE4UQkCJBkUh0lse-k3jl-cH2BPZtUvAkPBleNiAqKlufzz2y7kfIcwZrBky-3q67Lto1B9asQZSseUBWDBRUnIF4SFYAICuu2Jcj8iSlLQBjQqrH5Ig3qm5Ey1fkx-UN0ojBZD-N6cbPtMP8DXGk12FnzUhTjovNS8REzegofp_LNRWYBrzFkOjU_7wrHI5umoNJg7fFl71dwjJUhfUOx_zbNs1xyujHV_Tq3efqOix2SviGnv_zRA_pLuRoxtRjNAkpe0oe9SYkfHZ_HpOr92eXpx-qi0_nH09PLiorOM9VL63aqLYF4doNk65Bp2zbO5DCciM6qcAxibUywvCWSdaUPXQKLNYoLaj6mLw8eMtvvi6Ysh58shiCGXFakmayrWHTNs2moPyA2jilFLHXc_SDiTvNQO_b0Vu9b0fv29EgStaUoRf3_qUb0P0d-VNHAd4egLJYvPUYdbIeR4vOR7RZu8n_z_8LYnumKg</recordid><startdate>20150619</startdate><enddate>20150619</enddate><creator>Daikoku, Shusaku</creator><creator>Seko, Akira</creator><creator>Son, Sang-Hyun</creator><creator>Suzuki, Katsuhiko</creator><creator>Ito, Yukishige</creator><creator>Kanie, Osamu</creator><general>Elsevier Inc</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>7X8</scope><orcidid>https://orcid.org/0000-0003-4147-6868</orcidid></search><sort><creationdate>20150619</creationdate><title>The relationship between glycan structures and expression levels of an endoplasmic reticulum-resident glycoprotein, UDP-glucose: Glycoprotein glucosyltransferase 1</title><author>Daikoku, Shusaku ; Seko, Akira ; Son, Sang-Hyun ; Suzuki, Katsuhiko ; Ito, Yukishige ; Kanie, Osamu</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c422t-f6c9798804d8716d5ed9c8fd064c2a4b690d16e39a4a281615259b90ce3e6c093</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Chromatography, Liquid - methods</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Endoplasmic Reticulum - metabolism</topic><topic>Expression level</topic><topic>Glucosyltransferases - genetics</topic><topic>Glucosyltransferases - metabolism</topic><topic>Glycoproteins - genetics</topic><topic>Glycoproteins - metabolism</topic><topic>HEK293 Cells</topic><topic>Humans</topic><topic>Mass Spectrometry - methods</topic><topic>NanoLC–tandem MS</topic><topic>Polysaccharides - chemistry</topic><topic>Polysaccharides - metabolism</topic><topic>Recombinant Proteins - metabolism</topic><topic>Uridine 5′-diphosphate-glucose: glycoprotein glucosyltransferase 1</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Daikoku, Shusaku</creatorcontrib><creatorcontrib>Seko, Akira</creatorcontrib><creatorcontrib>Son, Sang-Hyun</creatorcontrib><creatorcontrib>Suzuki, Katsuhiko</creatorcontrib><creatorcontrib>Ito, Yukishige</creatorcontrib><creatorcontrib>Kanie, Osamu</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Biochemical and biophysical research communications</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Daikoku, Shusaku</au><au>Seko, Akira</au><au>Son, Sang-Hyun</au><au>Suzuki, Katsuhiko</au><au>Ito, Yukishige</au><au>Kanie, Osamu</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The relationship between glycan structures and expression levels of an endoplasmic reticulum-resident glycoprotein, UDP-glucose: Glycoprotein glucosyltransferase 1</atitle><jtitle>Biochemical and biophysical research communications</jtitle><addtitle>Biochem Biophys Res Commun</addtitle><date>2015-06-19</date><risdate>2015</risdate><volume>462</volume><issue>1</issue><spage>58</spage><epage>63</epage><pages>58-63</pages><issn>0006-291X</issn><eissn>1090-2104</eissn><abstract>In this article, we report a relationship between glycan structures and expression levels of a recombinant ER-resident glycoprotein, uridine 5′-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1). The function of glycan structures attached to a glycoprotein is actively studied; however, the glycan structures of recombinant, and not endogenous, glycoproteins have not been examined. In this study, we indicate a relationship between the glycan structure and the level of protein expression. Expression levels were controlled utilizing a series of vectors (pFN21K, pFN22K, pFN23K, and pFN24K HaloTag CMV Flexi Vectors). Qualitative and semi-quantitative confirmation of glycan structures was achieved with tandem mass spectrometry. The results of this study indicate that glycan structures are similar to endogenous glycans at low expression levels.
[Display omitted]
•The glycan profiles of glycoproteins were investigated using four expression vectors.•At low expression levels, glycan structures were similar to endogenous glycans.•The N-linked glycan structure of UGGT1 is dependent on the peptide expression level.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>25935482</pmid><doi>10.1016/j.bbrc.2015.04.105</doi><tpages>6</tpages><orcidid>https://orcid.org/0000-0003-4147-6868</orcidid></addata></record> |
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| subjects | Chromatography, Liquid - methods Electrophoresis, Polyacrylamide Gel Endoplasmic Reticulum - metabolism Expression level Glucosyltransferases - genetics Glucosyltransferases - metabolism Glycoproteins - genetics Glycoproteins - metabolism HEK293 Cells Humans Mass Spectrometry - methods NanoLC–tandem MS Polysaccharides - chemistry Polysaccharides - metabolism Recombinant Proteins - metabolism Uridine 5′-diphosphate-glucose: glycoprotein glucosyltransferase 1 |
| title | The relationship between glycan structures and expression levels of an endoplasmic reticulum-resident glycoprotein, UDP-glucose: Glycoprotein glucosyltransferase 1 |
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