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Phe31 acts as a gatekeeper for the substrate and product. (A) Phe31 (blue spheres) is fully in the χ1 = −60° rotamer when THF or FOL (yellow spheres) is bound, packing tightly against the substrate and pseudosubstrate. (B) In the hE:NADPH complex, with the vacant substrate binding pocket, Phe31 is fully in the χ1 = 180° rotamer, with the side chain packing against helix αC, leaving the substrate pocket accessible.
