Specific modification of a single cysteine residue in both bovine liver glutamate dehydrogenase and yeast glyceraldehyde-3-phosphate dehydrogenase. Difference in the mode of modification by pyrene maleimide.
Pyrene maleimide is shown to be a 'half of the sites' reagent for glutamate dehydrogenase and for glyceraldehyde-3-phosphate dehydrogenase. The modified residues are identified as cysteine-115 for glutamate dehydrogenase and cysteine-149 for glyceraldehyde-3-phosphate dehydrogenase. The two enzymes react differently with pyrene maleimide. Whereas the hydrophobic environment of cysteine-115 directs the modification of glutamate dehydrogenase, the high reactivity of cysteine-149 determines the specific modification of glyceraldehyde-3-phosphate dehydrogenase. Glutamate dehydrogenase activity is unaltered by the modification: glyceraldehyde-3-phosphate dehydrogenase activity in inhibited.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D003545:Cysteine; D005969:Glutamate Dehydrogenase / Q000378:metabolism*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008301:Maleimides / Q000494:pharmacology*; D010446:Peptide Fragments / Q000302:isolation & purification; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013439:Sulfhydryl Reagents / Q000494:pharmacology*; D014357:Trypsin
D000596:Amino Acids; D008301:Maleimides; D010446:Peptide Fragments; D013439:Sulfhydryl Reagents; C003944:N-(3-pyrene)maleimide; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D005969:Glutamate Dehydrogenase; D014357:Trypsin; D003545:Cysteine