) is a member of the family of heme
-containing peroxidases. Heme peroxidases
catalyse the H2O2-dependent oxidation of a wide range of different, usually organic, substrates in biology.
Ascorbate-dependent peroxidase activity was first reported in 1979,, more than 150 years after the first observation of peroxidase activity in horseradish plantsPlanche, LA. (1810) Bull. Pharm., 2, 578 and almost 40 years after the discovery of the closely related cytochrome c peroxidase
Peroxidases have been classified into three type (class I, class II and class III): ascorbate peroxidases is a class I peroxidase enzyme. APXs catalyse the H2O2-dependent oxidation of ascorbate
in plants, algae and certain cyanobacteria. APX has high sequence identity to cytochrome c peroxidase
, which is also a class I peroxidase enzyme. Under physiological conditions, the immediate product of the reaction, the monodehydroascorbate radical, is reduced back to ascorbate by a monodehydroascorbate reductase ( monodehydroascorbate reductase (NADH)
) enzyme. In the absence of a reductase, two monodehydroascorbate radicals disproportionate rapidly to dehydroascorbic acid and ascorbate
. APX is an integral component of the glutathione-ascorbate cycle
APX enzymes show high specificity for ascorbate as electron donor, but most APXs will also oxidise other organic substrates that are more characteristic of the class III peroxidases (such as horseradish peroxidase
), in some cases at rates comparable to that of ascorbate itself. This means that defining an enzyme as an APX is not straightforward, but is usually applied when the specific activity for ascorbate is higher than that for other substrates.
Most of the information on mechanism comes from work on the pea cytosolic and soybean cytosolic enzymes. The mechanism of oxidation of ascorbate is achieved by means of an oxidized Compound I intermediate, which is subsequently reduced by substrate in two, sequential single electron transfer steps (equations 1
, where HS = substrate and S• = one electron oxidised form of substrate).
APX + H2O2 → Compound I + H2O 1
Compound I + HS → Compound II + S• 2
Compound II + HS → APX + S• + H2O 3
In ascorbate peroxidase, Compound I is a transient (green) species and contains a high-valent iron
species (known as ferryl heme, FeIV) and a porphyrin
pi-cation radical,, as found in horseradish peroxidase. Compound II contains only the ferryl heme.
The structure of pea cytosolic APX was reported in 1995. The binding interaction of soybean cytosolic APX with its physiological substrate, ascorbate, and with a number of other substrates are also known.
Applications in cellular imaging
Both pea APX and soybean APX have been used in electron microscopy studies for cellular imaging.