7PGJ
Chimeric carminomycin-4-O-methyltransferase (DnrK) with regions from 10-decarboxylate TamK and 10-hydroxylase RdmB, together with a single point mutation F297G
Summary for 7PGJ
| Entry DOI | 10.2210/pdb7pgj/pdb |
| Descriptor | Carminomycin 4-O-methyltransferase DnrK,Methyltransferase domain-containing protein,Aclacinomycin 10-hydroxylase RdmB, S-ADENOSYL-L-HOMOCYSTEINE, methyl (1R,2R,4S)-2-ethyl-7-methoxy-2,4,5-tris(oxidanyl)-6,11-bis(oxidanylidene)-3,4-dihydro-1H-tetracene-1-carboxylate, ... (4 entities in total) |
| Functional Keywords | carminomycin-4-o-methyltransferase variant; chimeragenesis; aclacinomycin 10-hydroxylase, tsukubarubicin 10-decarboxylase; biosynthetic protein, biosynthetic protein |
| Biological source | Streptomyces peucetius More |
| Total number of polymer chains | 1 |
| Total formula weight | 40721.15 |
| Authors | Dinis, P.,MetsaKetela, M. (deposition date: 2021-08-14, release date: 2022-08-24, Last modification date: 2024-02-07) |
| Primary citation | Dinis, P.,Tirkkonen, H.,Wandi, B.N.,Siitonen, V.,Niemi, J.,Grocholski, T.,Metsa-Ketela, M. Evolution-inspired engineering of anthracycline methyltransferases. Pnas Nexus, 2:pgad009-pgad009, 2023 Cited by PubMed Abstract: soil bacteria produce hundreds of anthracycline anticancer agents with a relatively conserved set of genes. This diversity depends on the rapid evolution of biosynthetic enzymes to acquire novel functionalities. Previous work has identified -adenosyl-l-methionine-dependent methyltransferase-like proteins that catalyze 4-O-methylation, 10-decarboxylation, or 10-hydroxylation, with additional differences in substrate specificities. Here we focused on four protein regions to generate chimeric enzymes using sequences from four distinct subfamilies to elucidate their influence in catalysis. Combined with structural studies we managed to depict factors that influence gain-of-hydroxylation, loss-of-methylation, and substrate selection. The engineering expanded the catalytic repertoire to include novel 9,10-elimination activity, and 4-O-methylation and 10-decarboxylation of unnatural substrates. The work provides an instructive account on how the rise of diversity of microbial natural products may occur through subtle changes in biosynthetic enzymes. PubMed: 36874276DOI: 10.1093/pnasnexus/pgad009 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.13 Å) |
Structure validation
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