5n5d | pdb_00005n5d

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

spinqualitylabelsall models 5n5d, resolution 1.55Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 5n5d is a 2 chain structure with sequence from Streptomyces regensis. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 1.55Å Ligands: BU3, GOL, SAM, SO4 Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Publication Abstract from PubMed 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. Total Biosynthesis of the Pyrrolo[4,2]benzodiazepine Scaffold Tomaymycin on an In Vitro Reconstituted NRPS System.,von Tesmar A, Hoffmann M, Pippel J, Fayad AA, Dausend-Werner S, Bauer A, Blankenfeldt W, Muller R Cell Chem Biol. 2017 Aug 24. pii: S2451-9456(17)30275-1. doi:, 10.1016/j.chembiol.2017.08.001. PMID:28890318 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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