Structural highlights
Function
[PHEA_ECOLI] Catalyzes the Claisen rearrangement of chorismate to prephenate and the decarboxylation/dehydration of prephenate to phenylpyruvate.[1]
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[2]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Dopheide TA, Crewther P, Davidson BE. Chorismate mutase-prephenate dehydratase from Escherichia coli K-12. II. Kinetic properties. J Biol Chem. 1972 Jul 25;247(14):4447-52. PMID:4261395
- ↑ Koh M, Nasertorabi F, Han GW, Stevens RC, Schultz PG. Generation of an Orthogonal Protein-Protein Interface with a Noncanonical Amino Acid. J Am Chem Soc. 2017 Apr 26;139(16):5728-5731. doi: 10.1021/jacs.7b02273. Epub, 2017 Apr 17. PMID:28413876 doi:http://dx.doi.org/10.1021/jacs.7b02273
