4.1.3.16 hamging drop vapor diffusion method, using 100 mM HEPES-NaOH pH 7.0, 200 mM Na2SO4, 22.5% (w/v) PEG 3350, and 20% (v/v) glycerol or 100 mM Tris-HCl pH 8.0 and 1.8 M ammonium sulfate 4.1.3.16 to 1.97 A resolution, crystal structure of enzyme bound to pyruvate. Modeling of the 4-hydroxy-2-oxoglutarate-Schiff base intermediate and kinetic analyses of site-directed mutants support the importance of Lys196 as the nucleophile, Tyr168 and Ser77 as components of a proton relay, and Asn78 and Ser198 as unique residues that facilitate substrate binding