Understanding the enzymatic activity of 4-oxalocrotonate tautomerase and its mutant analogues: a computational study.
The effect of replacing arginine residues (Arg) with citrulline residues (Cit) in the binding site of 4-oxalocrotonate tautomerase (4-OT) was investigated with force field molecular dynamics and hybrid quantum mechanics/molecular mechanics studies. It is found that the Arg61Cit mutation has only minor effects on the k(cat) and K(M) values determined experimentally because of the flexibility of this residue. The decrease in k(cat) and increase in K(M) for the Arg11Cit and Arg39Cit mutations are due to the disruption of the binding site, which arises from repulsive interactions with neighboring residues. The results of this investigation shed new light on the effects of mutations in the binding site of 4-OT and consequently on how the enzyme binds the active substrate.
Tuttle
Tell
T
Max-Planck-Institut für Kohlenforschung, D-45470 Mülheim an der Ruhr, Germany. thiel@mpi-muelheim.mpg.de
Keinan
Ehud
E
Thiel
Walter
W
D001120:Arginine / Q000737:chemistry; D001665:Binding Sites; D002956:Citrulline / Q000737:chemistry; D003198:Computer Simulation; D030562:Databases, Protein; D019281:Dimerization; D004798:Enzymes / Q000737:chemistry*; D006860:Hydrogen Bonding; D007535:Isomerases / Q000737:chemistry*; D007700:Kinetics; D008956:Models, Chemical; D008968:Molecular Conformation; D009154:Mutation; D016958:Pseudomonas putida / Q000201:enzymology; D012984:Software
D004798:Enzymes; D002956:Citrulline; D001120:Arginine; C021918:4-oxalocrotonate tautomerase; D007535:Isomerases