An essential arginine residue at the substrate binding site of 4-hydroxyisophthalate hydroxylase.
4-Hydroxyisophthalate hydroxylase was inactivated by treatment with phenylglyoxal by a process obeying pseudo-first order kinetics indicating the presence of an essential arginine located presumably in the active site. Addition of saturating amounts of 4-hydroxyisophthalate during the treatment resulted in complete protection of the enzyme from the inactivation, but addition of NADPH was totally ineffective. Analysis of the effect of various substrate analogs on the protection of the enzyme showed that carboxyl and hydroxyl groups at para positions on the aromatic ring are essential for substrate binding to the active site. It was also observed that analogs which protect the enzyme against phenylglyoxal inactivation are themselves effective inhibitors of the enzyme activity.
D001120:Arginine; D001665:Binding Sites; D007700:Kinetics; D008024:Ligands; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D010658:Phenylglyoxal / Q000494:pharmacology; D011485:Protein Binding; D011549:Pseudomonas / Q000201:enzymology; D013329:Structure-Activity Relationship
D008024:Ligands; D001120:Arginine; D006899:Mixed Function Oxygenases; C021399:4-hydroxyisophthalate hydroxylase; D010658:Phenylglyoxal