7OQ6

Crystal structure of cytochrome P450 Sas16 from Streptomyces asterosporus

Summary for 7OQ6

• Entry DOI: 10.2210/pdb7oq6/pdb
• Descriptor: Cytochrome P450, PROTOPORPHYRIN IX CONTAINING FE, THIOCYANATE ION, ... (4 entities in total)
• Functional Keywords: cytochrome p450 monooxygenase, oxidoreductase
• Biological source: Streptomyces asterosporus
• Total number of polymer chains: 2
• Total formula weight: 100452.86

Authors

Zhang, L.,Zhang, S.,Bechthold, A.,Einsle, O. (deposition date: 2021-06-02, release date: 2022-06-22, Last modification date: 2024-11-06)

Primary citation

Zhang, S.,Zhang, L.,Greule, A.,Tailhades, J.,Marschall, E.,Prasongpholchai, P.,Leng, D.J.,Zhang, J.,Zhu, J.,Kaczmarski, J.A.,Schittenhelm, R.B.,Einsle, O.,Jackson, C.J.,Alberti, F.,Bechthold, A.,Zhang, Y.,Tosin, M.,Si, T.,Cryle, M.J.
P450-mediated dehydrotyrosine formation during WS9326 biosynthesis proceeds via dehydrogenation of a specific acylated dipeptide substrate.
Acta Pharm Sin B, 13:3561-3574, 2023

PubMed Abstract: WS9326A is a peptide antibiotic containing a highly unusual -methyl--2-3-dehydrotyrosine (NMet-Dht) residue that is incorporated during peptide assembly on a non-ribosomal peptide synthetase (NRPS). The cytochrome P450 encoded by (P450) has been shown to be essential for the formation of the alkene moiety in NMet-Dht, but the timing and mechanism of the P450-mediated ,-dehydrogenation of Dht remained unclear. Here, we show that the substrate of P450 is the NRPS-associated peptidyl carrier protein (PCP)-bound dipeptide intermediate ()-2-pent-1'-enyl-cinnamoyl-Thr--Me-Tyr. We demonstrate that P450-mediated incorporation of the double bond follows -methylation of the Tyr by the methyl transferase domain found within the NRPS, and further that P450 appears to be specific for substrates containing the ()-2-pent-1'-enyl-cinnamoyl group. A crystal structure of P450 reveals differences between P450 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 P450 activity and WS9326A biosynthesis. Together, our results suggest that P450 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.

PubMed: 37655329
DOI: 10.1016/j.apsb.2023.03.021

Experimental method

X-RAY DIFFRACTION (2 Å)