Ascorbate peroxidase

Ascorbate-dependent peroxidase activity was first reported in 1979,^[1],[2] more than 150 years after the first observation of peroxidase activity in horseradish plants^[3] and almost 40 years after the discovery of the closely related cytochrome c peroxidase enzyme.^[4]

Peroxidases have been classified into three types (class I, class II and class III): ascorbate peroxidases is a class I peroxidase enzyme.^[5] APXs catalyse the H₂O₂-dependent oxidation of ascorbate in plants, algae and certain cyanobacteria.^[6] 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.^[7]

APX enzymes show high specificity for ascorbate as an 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. Some proteins from the APX family lack the ascorbate-binding amino acid residues suggesting that they might oxidize other molecules than ascorbate.^[8]

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]–[3], 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, Fe^IV) and a porphyrin pi-cation radical,^[9],[10] as found in horseradish peroxidase. Compound II contains only the ferryl heme.

The structure of pea cytosolic APX was reported in 1995.^[11] The binding interaction of soybean cytosolic APX with its physiological substrate, ascorbate^[12],[13] and with a number of other substrates^[14] are also known.

As of late 2007, 12 structures have been solved for this class of enzymes, with PDB accession codes 1APX, 1IYN, 1OAF, 1OAG, 1V0H, 2CL4, 2GGN, 2GHC, 2GHD, 2GHE, 2GHH, and 2GHK.

Both pea APX^[15] and soybean APX and their mutants (APEX, APEX2)^[16] have been used in electron microscopy studies for cellular imaging.

• Cytochrome c peroxidase
• Manganese peroxidase

• EC 1.11.1.11 Archived 2011-05-16 at the Wayback Machine
• L-ascorbate peroxidase (EC-Number 1.11.1.11 ) Archived 2012-02-04 at the Wayback Machine