A convenient quantitative synthesis of methylglyoxal for glyoxalase I assays

Kellum, M.W.; Oray, Bedii; Norton, Scott J.

doi:10.1016/0003-2697(78)90258-0

Cited by 60 publications

(42 citation statements)

References 13 publications

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“…(8). Methylglyoxal was made by acid hydrolysis of pyru-The samples were derivatized at 20ЊC for 3.5 to 4 h. vic aldehyde dimethylacetal (Aldrich Chemical ComMedium sample preparation for extracellular methpany, Inc., Milwaukee, WI) as previously described (9). ylglyoxal measurements.…”

Section: Methodsmentioning

confidence: 99%

Method for Determination of Free Intracellular and Extracellular Methylglyoxal in Animal Cells Grown in Culture

Chaplen

Fahl

Cameron

1996

Analytical Biochemistry

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Section: Methodsmentioning

confidence: 99%

Method for Determination of Free Intracellular and Extracellular Methylglyoxal in Animal Cells Grown in Culture

Chaplen

Fahl

Cameron

1996

Analytical Biochemistry

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“…Meth-Series 1050 system (Hewlett-Packard Inc., U.S.A.). ylglyoxal was produced by acid hydrolysis of pyruvic The Hewlett-Packard system consisted of an autosamaldehyde dimethylacetal (Aldrich Chemical Co., Inc., pler, quaternary pump, and multidiode array detector Milwaukee, WI) as previously described (24). A D,L-and utilized HP3D ChemStations software.…”

Section: Methodsmentioning

confidence: 99%

Detection of Methylglyoxal as a Degradation Product of DNA and Nucleic Acid Components Treated with Strong Acid

Chaplen

Fahl

Cameron

1996

Analytical Biochemistry

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“…Glutathione and methylglyoxal dimethyl acetal were obtained from Sigma. Methylglyoxal was obtained by boiling of methylglyoxal dimethyl acetal for 10 min in 10% sulfuric acid (13). The glutathione derivative S-p-nitrocarbobenzoxyglutathione was a generous gift from Professor Giovanni Principato, Universita' di Ancona, Ancona, Italy.…”

Section: Methodsmentioning

confidence: 99%

Involvement of an Active-site Zn2+ Ligand in the Catalytic Mechanism of Human Glyoxalase I

Ridderström¹,

Cameron²,

Jones³

et al. 1998

Journal of Biological Chemistry

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The Zn 2؉ ligands glutamate 99 and glutamate 172 in the active site of human glyoxalase I were replaced, each in turn, by glutamines by site-directed mutagenesis to elucidate their potential significance for the catalytic properties of the enzyme. To compensate for the loss of the charged amino acid residue, another of the metal ligands, glutamine 33, was simultaneously mutated into glutamate. The double mutants and the single mutants Q33E, E99Q, and E172Q were expressed in Escherichia coli, purified on an S-hexylglutathione matrix, and characterized. Metal analysis demonstrated that mutant Q33E/E172Q contained 1.0 mol of zinc/mol of enzyme subunit, whereas mutant Q33E/E99Q contained only 0.3 mol of zinc/mol of subunit. No catalytic activity could be detected with the double mutant Q33E/E172Q (<10 ؊8 of the wild-type activity). The second double mutant Q33E/E99Q had 1.5% of the specific activity of the wild-type enzyme, whereas the values for mutants Q33E and E99Q were 1.3 and 0.1%, respectively; the E172Q mutant had less than 10 ؊5 times the specific activity of the wild-type.The crystal structure of the catalytically inactive double mutant Q33E/E172Q demonstrated that Zn 2؉ was bound without any gross changes or perturbations. The results suggest that the metal ligand glutamate 172 is directly involved in the catalytic mechanism of the enzyme, presumably serving as the base that abstracts a proton from the hemithioacetal substrate.

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A convenient quantitative synthesis of methylglyoxal for glyoxalase I assays

Cited by 60 publications

References 13 publications

Method for Determination of Free Intracellular and Extracellular Methylglyoxal in Animal Cells Grown in Culture

Method for Determination of Free Intracellular and Extracellular Methylglyoxal in Animal Cells Grown in Culture

Detection of Methylglyoxal as a Degradation Product of DNA and Nucleic Acid Components Treated with Strong Acid

Involvement of an Active-site Zn2+ Ligand in the Catalytic Mechanism of Human Glyoxalase I

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