Crystal structures of the most interesting VP/MnP isoenzymes from the P. ostreatus genome (1.05 to 1.10 Å). (A) Superposition of the overall structures of VP1 (light blue) and MnP4 (light brown) from two different orientations. The significant differences between the structures are shown in darker color with some side chains of these regions as sticks. (B) Electrostatic surface of VP1 and MnP4 from two different orientations showing the main heme access channel (larger circle) and the narrower Mn2+ access channel (smaller circle) as well as the more basic MnP4 surface with a high number of exposed lysine residues. (C) Detail of the VP1 and MnP4 heme and neighbor regions showing: proximal histidine (H169 and H175, respectively); distal histidine (H47) and neighbor arginine (R43); two conserved phenylalanines at the proximal histidine side (F186 and F182, respectively) and the distal histidine side (F46) of the heme; two conserved asparagines (N78 and N84, respectively) and glutamate residues (E72 and E78, respectively) forming a H-bond network from the distal histidine; two structural Ca2+ ions (brown spheres) with their four/five conserved ligands; one site for oxidation of Mn2+ (whose predicted position is marked with a red circle) near the internal propionate of heme, formed by two glutamates (E36 and E40) and one aspartate (D175 and D181, respectively); one tryptophan residue (W164) constituting the site (marked with a red benzenic ring) of VP1 oxidation of aromatic substrates by LRET via a contiguous leucine residue (L165); and several water molecules (white spheres) near the distal histidine and Ca2+ ion. From entries [PDB:4BLK] (VP1) and [PDB:4BM1] (MnP4). LRET, long-range electron transfer; MnP, manganese peroxidase; PDB, Protein Data Bank; VP, versatile peroxidase.