Post-translational site-selective protein backbone α-deuteration
Isotopic replacement has long-proven applications in small molecules. However, applications in proteins are largely limited to biosynthetic strategies or exchangeable (for example, N-H/D) labile sites only. The development of postbiosynthetic, C-1H → C-2H/D replacement in proteins could enable probi...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
Springer Nature
2018
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| _version_ | 1826266748171583488 |
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| author | Galan, S Wickens, J Dadova, J Ng, W Zhang, X Simion, R Quinlan, R Pires, E Paton, R Caddick, S Chudasama, V Davis, B |
| author_facet | Galan, S Wickens, J Dadova, J Ng, W Zhang, X Simion, R Quinlan, R Pires, E Paton, R Caddick, S Chudasama, V Davis, B |
| author_sort | Galan, S |
| collection | OXFORD |
| description | Isotopic replacement has long-proven applications in small molecules. However, applications in proteins are largely limited to biosynthetic strategies or exchangeable (for example, N-H/D) labile sites only. The development of postbiosynthetic, C-1H → C-2H/D replacement in proteins could enable probing of mechanisms, among other uses. Here we describe a chemical method for selective protein α-carbon deuteration (proceeding from Cys to dehydroalanine (Dha) to deutero-Cys) allowing overall 1H→2H/D exchange at a nonexchangeable backbone site. It is used here to probe mechanisms of reactions used in protein bioconjugation. This analysis suggests, together with quantum mechanical calculations, stepwise deprotonations via on-protein carbanions and unexpected sulfonium ylides in the conversion of Cys to Dha, consistent with a 'carba-Swern' mechanism. The ready application on existing, intact protein constructs (without specialized culture or genetic methods) suggests this C-D labeling strategy as a possible tool in protein mechanism, structure, biotechnology and medicine. |
| first_indexed | 2024-03-06T20:43:40Z |
| format | Journal article |
| id | oxford-uuid:351ea6db-cae9-4cb8-b1f3-674a10f495cb |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T20:43:40Z |
| publishDate | 2018 |
| publisher | Springer Nature |
| record_format | dspace |
| spelling | oxford-uuid:351ea6db-cae9-4cb8-b1f3-674a10f495cb2022-03-26T13:30:11ZPost-translational site-selective protein backbone α-deuterationJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:351ea6db-cae9-4cb8-b1f3-674a10f495cbEnglishSymplectic Elements at OxfordSpringer Nature2018Galan, SWickens, JDadova, JNg, WZhang, XSimion, RQuinlan, RPires, EPaton, RCaddick, SChudasama, VDavis, BIsotopic replacement has long-proven applications in small molecules. However, applications in proteins are largely limited to biosynthetic strategies or exchangeable (for example, N-H/D) labile sites only. The development of postbiosynthetic, C-1H → C-2H/D replacement in proteins could enable probing of mechanisms, among other uses. Here we describe a chemical method for selective protein α-carbon deuteration (proceeding from Cys to dehydroalanine (Dha) to deutero-Cys) allowing overall 1H→2H/D exchange at a nonexchangeable backbone site. It is used here to probe mechanisms of reactions used in protein bioconjugation. This analysis suggests, together with quantum mechanical calculations, stepwise deprotonations via on-protein carbanions and unexpected sulfonium ylides in the conversion of Cys to Dha, consistent with a 'carba-Swern' mechanism. The ready application on existing, intact protein constructs (without specialized culture or genetic methods) suggests this C-D labeling strategy as a possible tool in protein mechanism, structure, biotechnology and medicine. |
| spellingShingle | Galan, S Wickens, J Dadova, J Ng, W Zhang, X Simion, R Quinlan, R Pires, E Paton, R Caddick, S Chudasama, V Davis, B Post-translational site-selective protein backbone α-deuteration |
| title | Post-translational site-selective protein backbone α-deuteration |
| title_full | Post-translational site-selective protein backbone α-deuteration |
| title_fullStr | Post-translational site-selective protein backbone α-deuteration |
| title_full_unstemmed | Post-translational site-selective protein backbone α-deuteration |
| title_short | Post-translational site-selective protein backbone α-deuteration |
| title_sort | post translational site selective protein backbone α deuteration |
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