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Full-Text Articles in Physics
Converting Glx2-1 Into An Active Glyoxalase Ii, Pattraranee Limphong, Nicole E. Adams, Matthew F. Rouhier, Ross M. Mckinney, Melissa Naylor, Brian Bennett, Christopher A. Makaroff, Michael W. Crowder
Converting Glx2-1 Into An Active Glyoxalase Ii, Pattraranee Limphong, Nicole E. Adams, Matthew F. Rouhier, Ross M. Mckinney, Melissa Naylor, Brian Bennett, Christopher A. Makaroff, Michael W. Crowder
Physics Faculty Research and Publications
Arabidopsis thaliana glyoxalase 2-1 (GLX2-1) exhibits extensive sequence similarity with GLX2 enzymes but is catalytically inactive with SLG, the GLX2 substrate. In an effort to identify residues essential for GLX2 activity, amino acid residues were altered at positions 219, 246, 248, 325, and 328 in GLX2-1 to be the same as those in catalytically active human GLX2. The resulting enzymes were overexpressed, purified, and characterized using metal analyses, fluorescence spectroscopy, and steady-state kinetics to evaluate how these residues affect metal binding, structure, and catalysis. The R246H/N248Y double mutant exhibited low level S-lactoylglutathione hydrolase activity, while the R246H/N248Y/Q325R/R328K mutant exhibited …