Effect of pH on tobacco anionic peroxidase stability and its interaction with hydrogen peroxide.
The effect of extremely acidic pH on the stability of tobacco peroxidase and lignin peroxidase holoenzymes has been studied. Stabilization of tobacco peroxidase holoenzyme in the presence of calcium cations at pH < 2 and stabilization of lignin peroxidase at pH > 2 in the presence of veratryl alcohol have been shown. The dependence of the reaction rate constant for hydrogen peroxide interaction with tobacco peroxidase on pH suggests that the reaction rate is under control of a group with pK of 2.5. A tobacco peroxidase model structure has been created by means of homology modeling on the basis of the tobacco peroxidase sequence and the coordinates of peanut peroxidase crystal structure. The model structure demonstrates the presence of the negatively charged Glu-141 at the entrance to the active site and its electrostatic repulsion from heme propionates and triad of Asp-76, -79, and -80 residues. The results on tobacco holoperoxidase stabilization at pH 1.8 in the presence of calcium cations and drop in reaction rate constant for the enzyme interaction with hydrogen peroxide are explained by a hypothetical formation of ionic bonds between Glu-141 and the triad of aspartic acid residues via calcium cation lowering the accessibility of the active site and stabilizing the holoenzyme.
Gazaryan
I G
IG
Department of Chemical Enzymology, School of Chemistry, Lomonosov Moscow State University, Moscow, 119899, Russia.
Ouporov
I V
IV
Chubar
T A
TA
Fechina
V A
VA
Mareeva
E A
EA
Lagrimini
L M
LM
D000595:Amino Acid Sequence; D010367:Arachis / Q000201:enzymology / Q000235:genetics; D002118:Calcium / Q000378:metabolism; D020134:Catalytic Domain; D004795:Enzyme Stability; D020035:Holoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009195:Peroxidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010544:Peroxidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010947:Plants, Toxic; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D055672:Static Electricity; D014026:Nicotiana / Q000201:enzymology* / Q000235:genetics
D020035:Holoenzymes; D006861:Hydrogen Peroxide; D010544:Peroxidases; C042858:lignin peroxidase; D009195:Peroxidase; D002118:Calcium