5vht

E. coli chorismate mutase with orthogonal interface containing p-benzoyl phenylalanine

spinqualitylabelsall models 5vht, resolution 2.00Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 5vht is a 2 chain structure with sequence from Escherichia coli K-12. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 2Å Ligands: PBF Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function CMPDT_ECOLI Catalyzes the Claisen rearrangement of chorismate to prephenate and the decarboxylation/dehydration of prephenate to phenylpyruvate. Publication Abstract from PubMed We have engineered the protein interface of the Escherichia coli chorismate mutase (EcCM) homodimer to be dependent on incorporation of a noncanonical amino acid (ncAA) at residue 72. The large hydrophobic amino acid p-benzoyl phenylalanine (pBzF) was substituted for Tyr72, which led to a catalytically inactive protein. A library of five residues (Leu25', Arg29', Leu76, Ile80' and Asp83') surrounding pBzF72 was generated and subjected to a growth based selection in a chorismate mutase deficient strain. An EcCM variant (Phe25', pBzF72, Thr76, Gly80' and Tyr83') forms a stable homodimer, has catalytic activity similar to the wild type enzyme, and unfolds with a Tm of 53 degrees C. The X-ray crystal structure reveals a pi-pi stacking and hydrogen bonding interactions that stabilize the new protein interface. The strategy described here should be useful for generating organisms that are dependent on the presence of a ncAA for growth. Generation of an Orthogonal Protein-Protein Interface with a Noncanonical Amino Acid.,Koh M, Nasertorabi F, Han GW, Stevens RC, Schultz PG J Am Chem Soc. 2017 Apr 26;139(16):5728-5731. doi: 10.1021/jacs.7b02273. Epub, 2017 Apr 17. PMID:28413876 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See Also 3D structures of chorismate mutase

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