Site-selective glycosylation of proteins: creating synthetic glycoproteins.
In higher organisms, the functions of many proteins are modulated by post-translational modifications (PTMs). Glycosylation is by far the most diverse of the PTM processes. Natural protein production methods typically produce PTM or glycoform mixtures within which function is difficult to dissect or...
| Príomhchruthaitheoirí: | , , , |
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| Formáid: | Journal article |
| Teanga: | English |
| Foilsithe / Cruthaithe: |
2007
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| _version_ | 1826273137472307200 |
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| author | van Kasteren, S Kramer, H Gamblin, D Davis, B |
| author_facet | van Kasteren, S Kramer, H Gamblin, D Davis, B |
| author_sort | van Kasteren, S |
| collection | OXFORD |
| description | In higher organisms, the functions of many proteins are modulated by post-translational modifications (PTMs). Glycosylation is by far the most diverse of the PTM processes. Natural protein production methods typically produce PTM or glycoform mixtures within which function is difficult to dissect or control. Chemical tagging methods allow the precise attachment of multiple glycosylation modifications to bacterially expressed (bare) protein scaffolds, allowing reconstitution of functionally effective mimics of glycoproteins in higher organisms. In this way combining chemical control of PTM with readily available protein scaffolds provides a systematic platform for creating probes of protein-PTM interactions. This protocol describes the modification of Cys residues in proteins using glycomethanethiosulfonates and glycoselenenylsulfides and the modification of azidohomoalanine residues, introduced by Met replacement using auxotrophic Met(-) Escherichia coli strains, with glycoalkynes and the combination of these techniques for the creation of dual-tagged proteins. Each glycosylation procedure outlined in this protocol can be achieved in half a day. |
| first_indexed | 2024-03-06T22:23:36Z |
| format | Journal article |
| id | oxford-uuid:55e380d6-f8c5-4218-aba8-436958d2277c |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T22:23:36Z |
| publishDate | 2007 |
| record_format | dspace |
| spelling | oxford-uuid:55e380d6-f8c5-4218-aba8-436958d2277c2022-03-26T16:47:03ZSite-selective glycosylation of proteins: creating synthetic glycoproteins.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:55e380d6-f8c5-4218-aba8-436958d2277cEnglishSymplectic Elements at Oxford2007van Kasteren, SKramer, HGamblin, DDavis, BIn higher organisms, the functions of many proteins are modulated by post-translational modifications (PTMs). Glycosylation is by far the most diverse of the PTM processes. Natural protein production methods typically produce PTM or glycoform mixtures within which function is difficult to dissect or control. Chemical tagging methods allow the precise attachment of multiple glycosylation modifications to bacterially expressed (bare) protein scaffolds, allowing reconstitution of functionally effective mimics of glycoproteins in higher organisms. In this way combining chemical control of PTM with readily available protein scaffolds provides a systematic platform for creating probes of protein-PTM interactions. This protocol describes the modification of Cys residues in proteins using glycomethanethiosulfonates and glycoselenenylsulfides and the modification of azidohomoalanine residues, introduced by Met replacement using auxotrophic Met(-) Escherichia coli strains, with glycoalkynes and the combination of these techniques for the creation of dual-tagged proteins. Each glycosylation procedure outlined in this protocol can be achieved in half a day. |
| spellingShingle | van Kasteren, S Kramer, H Gamblin, D Davis, B Site-selective glycosylation of proteins: creating synthetic glycoproteins. |
| title | Site-selective glycosylation of proteins: creating synthetic glycoproteins. |
| title_full | Site-selective glycosylation of proteins: creating synthetic glycoproteins. |
| title_fullStr | Site-selective glycosylation of proteins: creating synthetic glycoproteins. |
| title_full_unstemmed | Site-selective glycosylation of proteins: creating synthetic glycoproteins. |
| title_short | Site-selective glycosylation of proteins: creating synthetic glycoproteins. |
| title_sort | site selective glycosylation of proteins creating synthetic glycoproteins |
| work_keys_str_mv | AT vankasterens siteselectiveglycosylationofproteinscreatingsyntheticglycoproteins AT kramerh siteselectiveglycosylationofproteinscreatingsyntheticglycoproteins AT gamblind siteselectiveglycosylationofproteinscreatingsyntheticglycoproteins AT davisb siteselectiveglycosylationofproteinscreatingsyntheticglycoproteins |