P450-mediated dehydrotyrosine formation during WS9326 biosynthesis proceeds via dehydrogenation of a specific acylated dipeptide substrate
© 2023 Chinese Pharmaceutical Association and Institute of Materia Medica, Chinese Academy of Medical Sciences. Production and hosting by Elsevier B.V..
WS9326A is a peptide antibiotic containing a highly unusual N-methyl-E-2-3-dehydrotyrosine (NMet-Dht) residue that is incorporated during peptide assembly on a non-ribosomal peptide synthetase (NRPS). The cytochrome P450 encoded by sas16 (P450Sas) has been shown to be essential for the formation of the alkene moiety in NMet-Dht, but the timing and mechanism of the P450Sas-mediated α,β-dehydrogenation of Dht remained unclear. Here, we show that the substrate of P450Sas is the NRPS-associated peptidyl carrier protein (PCP)-bound dipeptide intermediate (Z)-2-pent-1'-enyl-cinnamoyl-Thr-N-Me-Tyr. We demonstrate that P450Sas-mediated incorporation of the double bond follows N-methylation of the Tyr by the N-methyl transferase domain found within the NRPS, and further that P450Sas appears to be specific for substrates containing the (Z)-2-pent-1'-enyl-cinnamoyl group. A crystal structure of P450Sas reveals differences between P450Sas and other P450s involved in the modification of NRPS-associated substrates, including the substitution of the canonical active site alcohol residue with a phenylalanine (F250), which in turn is critical to P450Sas activity and WS9326A biosynthesis. Together, our results suggest that P450Sas catalyses the direct dehydrogenation of the NRPS-bound dipeptide substrate, thus expanding the repertoire of P450 enzymes that can be used to produce biologically active peptides.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:13 |
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| Enthalten in: |
Acta pharmaceutica Sinica. B - 13(2023), 8 vom: 02. Aug., Seite 3561-3574 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Zhang, Songya [VerfasserIn] |
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| Links: |
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| Themen: |
Cytochrome P450 |
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| Anmerkungen: |
Date Revised 02.09.2023 published: Print-Electronic Citation Status PubMed-not-MEDLINE |
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| doi: |
10.1016/j.apsb.2023.03.021 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM361531184 |
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| 520 | |a WS9326A is a peptide antibiotic containing a highly unusual N-methyl-E-2-3-dehydrotyrosine (NMet-Dht) residue that is incorporated during peptide assembly on a non-ribosomal peptide synthetase (NRPS). The cytochrome P450 encoded by sas16 (P450Sas) has been shown to be essential for the formation of the alkene moiety in NMet-Dht, but the timing and mechanism of the P450Sas-mediated α,β-dehydrogenation of Dht remained unclear. Here, we show that the substrate of P450Sas is the NRPS-associated peptidyl carrier protein (PCP)-bound dipeptide intermediate (Z)-2-pent-1'-enyl-cinnamoyl-Thr-N-Me-Tyr. We demonstrate that P450Sas-mediated incorporation of the double bond follows N-methylation of the Tyr by the N-methyl transferase domain found within the NRPS, and further that P450Sas appears to be specific for substrates containing the (Z)-2-pent-1'-enyl-cinnamoyl group. A crystal structure of P450Sas reveals differences between P450Sas and other P450s involved in the modification of NRPS-associated substrates, including the substitution of the canonical active site alcohol residue with a phenylalanine (F250), which in turn is critical to P450Sas activity and WS9326A biosynthesis. Together, our results suggest that P450Sas catalyses the direct dehydrogenation of the NRPS-bound dipeptide substrate, thus expanding the repertoire of P450 enzymes that can be used to produce biologically active peptides | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Cytochrome P450 | |
| 650 | 4 | |a Enzyme mechanism | |
| 650 | 4 | |a Natural products | |
| 650 | 4 | |a Non-ribosomal peptide synthetase | |
| 650 | 4 | |a Peptide antibiotic | |
| 650 | 4 | |a Protein crystal structure | |
| 700 | 1 | |a Zhang, Lin |e verfasserin |4 aut | |
| 700 | 1 | |a Greule, Anja |e verfasserin |4 aut | |
| 700 | 1 | |a Tailhades, Julien |e verfasserin |4 aut | |
| 700 | 1 | |a Marschall, Edward |e verfasserin |4 aut | |
| 700 | 1 | |a Prasongpholchai, Panward |e verfasserin |4 aut | |
| 700 | 1 | |a Leng, Daniel J |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Jingfan |e verfasserin |4 aut | |
| 700 | 1 | |a Zhu, Jing |e verfasserin |4 aut | |
| 700 | 1 | |a Kaczmarski, Joe A |e verfasserin |4 aut | |
| 700 | 1 | |a Schittenhelm, Ralf B |e verfasserin |4 aut | |
| 700 | 1 | |a Einsle, Oliver |e verfasserin |4 aut | |
| 700 | 1 | |a Jackson, Colin J |e verfasserin |4 aut | |
| 700 | 1 | |a Alberti, Fabrizio |e verfasserin |4 aut | |
| 700 | 1 | |a Bechthold, Andreas |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Youming |e verfasserin |4 aut | |
| 700 | 1 | |a Tosin, Manuela |e verfasserin |4 aut | |
| 700 | 1 | |a Si, Tong |e verfasserin |4 aut | |
| 700 | 1 | |a Cryle, Max J |e verfasserin |4 aut | |
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