Quantitative N- or C-Terminal Labelling of Proteins with Unactivated Peptides by Use of Sortases and a d-Aminopeptidase
© 2023 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH..
Quantitative and selective labelling of proteins is widely used in both academic and industrial laboratories, and catalytic labelling of proteins using transpeptidases, such as sortases, has proved to be a popular strategy for such selective modification. A major challenge for this class of enzymes is that the majority of procedures require an excess of the labelling reagent or, alternatively, activated substrates rather than simple commercially sourced peptides. We report the use of a coupled enzyme strategy which enables quantitative N- and C-terminal labelling of proteins using unactivated labelling peptides. The use of an aminopeptidase in conjunction with a transpeptidase allows sequence-specific degradation of the peptide by-product, shifting the equilibrium to favor product formation, which greatly enhances the reaction efficiency. Subsequent optimisation of the reaction allows N-terminal labelling of proteins using essentially equimolar ratios of peptide label to protein and C-terminal labelling with only a small excess. Minimizing the amount of substrate required for quantitative labelling has the potential to improve industrial processes and facilitate the use of transpeptidation as a method for protein labelling.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:63 |
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| Enthalten in: |
Angewandte Chemie (International ed. in English) - 63(2024), 8 vom: 19. Feb., Seite e202310862 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Arnott, Zoe L P [VerfasserIn] |
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| Links: |
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| Themen: |
Aminoacyltransferases |
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| Anmerkungen: |
Date Completed 14.02.2024 Date Revised 14.02.2024 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1002/anie.202310862 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 245 | 1 | 0 | |a Quantitative N- or C-Terminal Labelling of Proteins with Unactivated Peptides by Use of Sortases and a d-Aminopeptidase |
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| 520 | |a © 2023 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH. | ||
| 520 | |a Quantitative and selective labelling of proteins is widely used in both academic and industrial laboratories, and catalytic labelling of proteins using transpeptidases, such as sortases, has proved to be a popular strategy for such selective modification. A major challenge for this class of enzymes is that the majority of procedures require an excess of the labelling reagent or, alternatively, activated substrates rather than simple commercially sourced peptides. We report the use of a coupled enzyme strategy which enables quantitative N- and C-terminal labelling of proteins using unactivated labelling peptides. The use of an aminopeptidase in conjunction with a transpeptidase allows sequence-specific degradation of the peptide by-product, shifting the equilibrium to favor product formation, which greatly enhances the reaction efficiency. Subsequent optimisation of the reaction allows N-terminal labelling of proteins using essentially equimolar ratios of peptide label to protein and C-terminal labelling with only a small excess. Minimizing the amount of substrate required for quantitative labelling has the potential to improve industrial processes and facilitate the use of transpeptidation as a method for protein labelling | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Coupling Enzymes | |
| 650 | 4 | |a Protein Labelling | |
| 650 | 4 | |a Sortase | |
| 650 | 4 | |a Transpeptidase | |
| 650 | 7 | |a Aminopeptidases |2 NLM | |
| 650 | 7 | |a EC 3.4.11.- |2 NLM | |
| 650 | 7 | |a Bacterial Proteins |2 NLM | |
| 650 | 7 | |a Aminoacyltransferases |2 NLM | |
| 650 | 7 | |a EC 2.3.2.- |2 NLM | |
| 650 | 7 | |a Peptides |2 NLM | |
| 650 | 7 | |a Peptidyl Transferases |2 NLM | |
| 650 | 7 | |a EC 2.3.2.12 |2 NLM | |
| 700 | 1 | |a Morgan, Holly E |e verfasserin |4 aut | |
| 700 | 1 | |a Hollingsworth, Kristian |e verfasserin |4 aut | |
| 700 | 1 | |a Stevenson, Charlotte M E |e verfasserin |4 aut | |
| 700 | 1 | |a Collins, Lawrence J |e verfasserin |4 aut | |
| 700 | 1 | |a Tamasanu, Alexandra |e verfasserin |4 aut | |
| 700 | 1 | |a Machin, Darren C |e verfasserin |4 aut | |
| 700 | 1 | |a Dolan, Jonathan P |e verfasserin |4 aut | |
| 700 | 1 | |a Kamiński, Tomasz P |e verfasserin |4 aut | |
| 700 | 1 | |a Wildsmith, Gemma C |e verfasserin |4 aut | |
| 700 | 1 | |a Williamson, Daniel J |e verfasserin |4 aut | |
| 700 | 1 | |a Pickles, Isabelle B |e verfasserin |4 aut | |
| 700 | 1 | |a Warriner, Stuart L |e verfasserin |4 aut | |
| 700 | 1 | |a Turnbull, W Bruce |e verfasserin |4 aut | |
| 700 | 1 | |a Webb, Michael E |e verfasserin |4 aut | |
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