6xn6

ScoE with the CABA substrate bound and His299 and Arg157 flipped out

spinqualitylabelsall models 6xn6, resolution 1.70Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 6xn6 is a 1 chain structure with sequence from Streptomyces coeruleorubidus. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 1.7Å Ligands: 7UC, ACT, CL, FE2 Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function A0A3B6UEU3_STRC4 Publication Abstract from PubMed The isonitrile moiety is found in marine sponges and some microbes, where it plays a role in processes such as virulence and metal acquisition. Until recently only one route was known for isonitrile biosynthesis, a condensation reaction that brings together a nitrogen atom of l-Trp/l-Tyr with a carbon atom from ribulose-5-phosphate. With the discovery of ScoE, a mononuclear Fe(II) alpha-ketoglutarate-dependent dioxygenase from Streptomyces coeruleorubidus, a second route was identified. ScoE forms isonitrile from a glycine adduct, with both the nitrogen and carbon atoms coming from the same glycyl moiety. This reaction is part of the nonribosomal biosynthetic pathway of isonitrile lipopeptides. Here, we present structural, biochemical and computational investigations of the mechanism of isonitrile formation by ScoE, an unprecedented reaction in the mononuclear Fe(II) alpha-ketoglutarate-dependent dioxygenase superfamily. The stoichiometry of this enzymatic reaction is measured and multiple high-resolution (1.45-1.96 A resolution) crystal structures of Fe(II)-bound ScoE are presented, providing insight into the binding of substrate, (R)-3-((carboxylmethyl)amino)butanoic acid (CABA), co-substrate alpha-ketoglutarate, and an Fe(IV)=O mimic oxovanadium. Comparison to a previously published crystal structure of ScoE suggests that ScoE has an 'inducible' alpha-ketoglutarate binding site, in which two residues arginine-157 and histidine-299 move by approximately 10 A from the surface of the protein into the active site to create a transient alpha-ketoglutarate binding pocket. Together, data from structural analyses, site-directed mutagenesis and computation, provide insight into the mode of alpha-ketoglutarate binding, the mechanism of isonitrile formation, and how the structure of ScoE has been adapted to perform this unusual chemical reaction. Biochemical and crystallographic investigations into isonitrile formation by a non-heme iron-dependent oxidase/decarboxylase.,Jonnalagadda R, Del Rio Flores A, Cai W, Mehmood R, Narayanamoorthy M, Ren C, Zaragoza JPT, Kulik HJ, Zhang W, Drennan CL J Biol Chem. 2020 Dec 27. pii: RA120.015932. doi: 10.1074/jbc.RA120.015932. PMID:33361191 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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