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Significance of active site methionine. (A) Overlay of Rg3GUS (PDB 6MVG) and FpL2-6 (PDB 6MVF) the fastest and slowest GUS enzymes, respectively. (B) Inset of Rg3GUS and FpL2-6 GUS active sites and modeled regorafenib-glucuronide. Where Rg3GUS contains a methionine, FpL2-6 contains a glycine. Catalytic glutamates are highlighted in magenta. (C) Rates of FpL2-6 mutant G380M. Mutation to methionine increases the rate by 40-fold.
