5N5D

Crystal Structure of the O-Methyltransferase TomG from Streptomyces achromogenes involved in Tomaymycin synthesis in complex with SAM

Summary for 5N5D

• Entry DOI: 10.2210/pdb5n5d/pdb
• Descriptor: Methyltransferase, (R,R)-2,3-BUTANEDIOL, GLYCEROL, ... (6 entities in total)
• Functional Keywords: o-methyltransferase, tomaymycin, rossmann-like fold, s-adenosyl methionine, transferase
• Biological source: Streptomyces regensis
• Total number of polymer chains: 2
• Total formula weight: 51157.80

Authors

Pippel, J.,Blankenfeldt, W. (deposition date: 2017-02-13, release date: 2017-09-20, Last modification date: 2024-01-17)

Primary citation

von Tesmar, A.,Hoffmann, M.,Pippel, J.,Fayad, A.A.,Dausend-Werner, S.,Bauer, A.,Blankenfeldt, W.,Muller, R.
Total Biosynthesis of the Pyrrolo[4,2]benzodiazepine Scaffold Tomaymycin on an In Vitro Reconstituted NRPS System.
Cell Chem Biol, 24:1216-1227.e8, 2017

PubMed Abstract: In vitro reconstitution and biochemical analysis of natural product biosynthetic pathways remains a challenging endeavor, especially if megaenzymes of the nonribosomal peptide synthetase (NRPS) type are involved. In theory, all biosynthetic steps may be deciphered using mass spectrometry (MS)-based analyses of both the carrier protein-coupled intermediates and the free intermediates. We here report the "total biosynthesis" of the pyrrolo[4,2]benzodiazepine scaffold tomaymycin using an in vitro reconstituted NRPS system. Proteoforms were analyzed by liquid chromatography (LC)-MS to decipher every step of the biosynthesis on its respective megasynthetase with up to 170 kDa in size. To the best of our knowledge, this is the first report of a comprehensive analysis of virtually all chemical steps involved in the biosynthesis of nonribosomally synthesized natural products. The study includes experiments to determine substrate specificities of the corresponding A-domains in competition assays by analyzing the adenylation step as well as the transfer to the respective carrier protein domain.

PubMed: 28890318
DOI: 10.1016/j.chembiol.2017.08.001

Experimental method

X-RAY DIFFRACTION (1.55 Å)