5N81

Crystal structure of an engineered TycA variant in complex with an O-propargyl-beta-Tyr-AMP analog

Summary for 5N81

• Entry DOI: 10.2210/pdb5n81/pdb
• Descriptor: Tyrocidine synthase 1, [(2~{R},3~{S},4~{R},5~{R})-5-(6-aminopurin-9-yl)-3,4-bis(oxidanyl)oxolan-2-yl]methyl ~{N}-[(3~{S})-3-azanyl-3-(4-prop-2-ynoxyphenyl)propanoyl]sulfamate, SULFATE ION, ... (5 entities in total)
• Functional Keywords: nonribosomal peptide synthetase, adenylation domain, ligase
• Biological source: Brevibacillus parabrevis
• Total number of polymer chains: 2
• Total formula weight: 96870.49

Authors

Niquille, D.L.,Hansen, D.A.,Mori, T.,Fercher, D.,Kries, H.,Hilvert, D. (deposition date: 2017-02-22, release date: 2017-12-13, Last modification date: 2024-01-17)

Primary citation

Niquille, D.L.,Hansen, D.A.,Mori, T.,Fercher, D.,Kries, H.,Hilvert, D.
Nonribosomal biosynthesis of backbone-modified peptides.
Nat Chem, 10:282-287, 2018

PubMed Abstract: Biosynthetic modification of nonribosomal peptide backbones represents a potentially powerful strategy to modulate the structure and properties of an important class of therapeutics. Using a high-throughput assay for catalytic activity, we show here that an L-Phe-specific module of an archetypal nonribosomal peptide synthetase can be reprogrammed to accept and process the backbone-modified amino acid (S)-β-Phe with near-native specificity and efficiency. A co-crystal structure with a non-hydrolysable aminoacyl-AMP analogue reveals the origins of the 40,000-fold α/β-specificity switch, illuminating subtle but precise remodelling of the active site. When the engineered catalyst was paired with downstream module(s), (S)-β-Phe-containing peptides were produced at preparative scale in vitro (~1 mmol) and high titres in vivo (~100 mg l), highlighting the potential of biosynthetic pathway engineering for the construction of novel nonribosomal β-frameworks.

PubMed: 29461527
DOI: 10.1038/nchem.2891

Experimental method

X-RAY DIFFRACTION (1.6 Å)