Converting GLX2-1 into an active glyoxalase II

The enzymes obtained from the alteration of amino acids residues at positions 219, 246, 248, 325, and 328 in Arabidopsis thaliana glyoxalase 2-1 (GLX2-1) were overexpressed, purified, and characterized to identify residues essential for GLX2 substrate activity. The results revealed that an active GLX2 enzyme requires both the presence of a properly positioned metal center and significant nonmetal, enzyme-substrate contacts, with the most important tyrosine 255.