1.3.98.1 C130A low activity with dihydroorotate, increasing activity above pH 8.0 with dihydrooxonate 390913 1.3.98.1 C130A the mutant enzyme exhibits binding affinities for dihydroorotate similar to that of the wild type enzyme, reduction is extremely slow compared to that of the wild type, the rate of reduction increases with pH showing no sign of a plateau 685146 1.3.98.1 C130A the mutant forms charge-transfer complexes upon binding 3,4-dihydroxybenzoate, but the maximum of the broad charge-transfer bands is shifted to 590 nm 685158 1.3.98.1 C130S the mutant enzyme exhibits binding affinities for dihydroorotate similar to that of the wild type enzyme, reduction is extremely slow compared to that of the wild type, the rate of reduction increases with pH showing no sign of a plateau 685146 1.3.98.1 C130S the mutant forms charge-transfer complexes upon binding 3,4-dihydroxybenzoate, but the maximum of the broad charge-transfer bands is shifted to 610 nm 685158 1.3.98.1 E206/K296E conversion of intermolecular salt bridge, mutant is fully active in concentrated solutions and dissociates into monomers upon dilution like wild-type enzyme 657291 1.3.98.1 E206A disturbance of intermolecular salt bridge, mutant retains almost complete activity 657291 1.3.98.1 E206K disturbance of intermolecular salt bridge, mutant activity is severely impaired 657291 1.3.98.1 H185A 4fold increase in KM-value of CoQD, 50% increase in KM-value of L-dihydroorotate 656285 1.3.98.1 K136E little changes in activity 656148 1.3.98.1 K213E decrease in activity against dichlorophenolindophenol 656148 1.3.98.1 K296A disturbance of intermolecular salt bridge, mutant retains almost complete activity 657291 1.3.98.1 K296E disturbance of intermolecular salt bridge, mutant activity is severely impaired 657291 1.3.98.1 L71F reaction is somehow slower than for wild-type, binding of dihydroorotate is much tighter than with wild-type 724305 1.3.98.1 L71F/C130S/V133T addition of the two residues comprising the conserved proton-transfer network of Class 2 dihydroorotate dehydrogenase from Escherichia coli to the C130S Class 1A enzyme of Lactococcus lacits. Mutation does not did not restore the function of the active site base or rapid flavin reduction. Kd for dihydroorotate is about three times tighter than the wild-type 724305 1.3.98.1 L71F/V133T reaction is drastically slower forwith wild-type, Kd for dihydroorotate is iabout eight-fold tighter than the wild-type 724305 1.3.98.1 additional information deletions of N-terminal residues from 2 down to 40 result in generally unstable proteins with apo protein quickly precipitating and FMN remaining in solution. A truncated protein lacking residues 2 to 30 is sufficiently stable, has near to wild-type activity using molecular oxygen and 20fold lower activity using 2,6-dichlorophenolindophenol as electron acceptor 657355 1.3.98.1 additional information DHOD-knockout Trypanosoma cruzi do not express the enzyme protein and can not survive even in the presence of pyrimidine nucleosides, suggesting a vital role of fumarate reductase activity in the regulation of cellular redox balance 675478 1.3.98.1 N127A reduced activity 656148 1.3.98.1 N193A drastically reduced activiy 656148 1.3.98.1 N67A drastically reduced activiy 656148 1.3.98.1 P131A decrease in activtiy against fumarate 656148 1.3.98.1 P56A drastically reduced activiy 656148 1.3.98.1 Q7L single nucleotide polymorphism due to missense polymorphism 19C>A, leading to amino acid substitution in the cationic N-terminal region of the polypeptide 700606 1.3.98.1 R265A 2fold increase in KM-value of CoQD, 15% increase in KM-value of L-dihydroorotate 656285 1.3.98.1 R50E little changes in activity 656148 1.3.98.1 R57A increase in activity 656148 1.3.98.1 S129A decrease in activtiy against fumarate 656148 1.3.98.1 S175A - 390913 1.3.98.1 S175A low activity with dihydroorotate, increasing activity above pH 8.0 with dihydrooxonate 390913 1.3.98.1 V133T reaction is somehow slower than for wild-type, binding of dihydroorotate is much weaker than with wild-type 724305