The crystal structure of peanut peroxidase

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The crystal structure of peanut peroxidase David J Schuller, Nenad Ban, Robert B van Huystee, Alexander McPherson, Thomas L Poulos  Structure  Volume 4, Issue 3, Pages 311-321 (March 1996) DOI: 10.1016/S0969-2126(96)00035-4

Figure 1 Stereo Cα trace of PNP molecule A, with every 20th residue labeled. The calcium atoms and potential glycosylation sites Asn60, Asn144 and Asn185 are also labeled. (Figure prepared with SETOR [56].) Structure 1996 4, 311-321DOI: (10.1016/S0969-2126(96)00035-4)

Figure 2 Ribbon diagrams of yeast cytochrome c peroxidase (CCP), pea cytosolic ascorbate peroxidase (APX), lignin peroxidase from P. chrysosporium (LIP) and peanut peroxidase (PNP) with helices labeled. Conserved helices appear in blue and the β-sheet of CCP in violet. The hemes are displayed in red and the proximal histidine ligands in pink. Disulfide bonds appear in yellow. Bound cations are shown as orange spheres and the helices unique to PNP are highlighted in green. (Figure prepared with SETOR[56].) Structure 1996 4, 311-321DOI: (10.1016/S0969-2126(96)00035-4)

Figure 2 Ribbon diagrams of yeast cytochrome c peroxidase (CCP), pea cytosolic ascorbate peroxidase (APX), lignin peroxidase from P. chrysosporium (LIP) and peanut peroxidase (PNP) with helices labeled. Conserved helices appear in blue and the β-sheet of CCP in violet. The hemes are displayed in red and the proximal histidine ligands in pink. Disulfide bonds appear in yellow. Bound cations are shown as orange spheres and the helices unique to PNP are highlighted in green. (Figure prepared with SETOR[56].) Structure 1996 4, 311-321DOI: (10.1016/S0969-2126(96)00035-4)

Figure 3 Alignment of CCP, APX, LIP and PNP sequences based on a structural superposition, with the sequence of HRP C added based on an alignment of HRP C and PNP. The secondary structure of PNP is indicated in the shaded row labeled ‘PNP 2D’. Residues for which the Cα coordinates superimpose with a root mean square deviation of <6.0 å are displayed in bold upper-case characters and are boxed. The distal and proximal heme pocket residues of all the proteins are shaded. Cysteine residues of PNP and HRP C are highlighted in inverse text with the disulfide pairings indicated by numerals. The ‘z’ at the N terminus of PNP and HRP C indicates a PCA residue. Calcium ligands of PNP are labeled with ‘+’. Carbohydrate attachment sites of PNP are labelled with ‘&’. The heterogeneous C terminus of mature HRP C is indicated with triangles [18]. cDNA sequences for HRP C (OPRHC) [39] and PNP (A38265) [16] were obtained from the PIR database. Coordinates for CCP (2CYP) [6] and LIP (1LGA) [8] were obtained from the Protein Data Bank [55]. The APX coordinates (1APX) [7] were kindly provided by William R Patterson. The alignment was created with the aid of STAMP [57], AMPS [58] and ALSCRIPT [59]. Structure 1996 4, 311-321DOI: (10.1016/S0969-2126(96)00035-4)

Figure 4 The heme environment of PNP. In this view, the distal heme pocket is above the plane of the heme and the proximal pocket is below it. The polypeptide and heme are green, solvent atoms are blue, and the 2Fo–Fc electron-density map is red. Surrounding residues have been removed to improve clarity. The map is contoured at 1σ. (Figure prepared with SETOR [56].) Structure 1996 4, 311-321DOI: (10.1016/S0969-2126(96)00035-4)

Figure 5 The substrate access channel of PNP. Side chains are colored by residue type. Residues behind the heme are clipped from view. The glycosylation site of Asn144 is visible in the lower left corner, but no carbohydrate is shown. (Figure prepared with SETOR [56].) Structure 1996 4, 311-321DOI: (10.1016/S0969-2126(96)00035-4)

Figure 6 Calcium sites. (a) Distal calcium site of PNP with the equivalent regions of CCP, APX and LIP superimposed. Colors are: CCP red, APX green, LIP purple, and PNP gold. (b) Distal calcium site of PNP in stereo with calcium ligands indicated by dotted lines. (c) Proximal calcium site of PNP with the equivalent regions of CCP, APX and LIP superimposed. Colors are the same as in (a). (d) Proximal calcium site of PNP in stereo with calcium ligands indicated by dotted lines. (Figure prepared with SETOR[56].) Structure 1996 4, 311-321DOI: (10.1016/S0969-2126(96)00035-4)

Figure 6 Calcium sites. (a) Distal calcium site of PNP with the equivalent regions of CCP, APX and LIP superimposed. Colors are: CCP red, APX green, LIP purple, and PNP gold. (b) Distal calcium site of PNP in stereo with calcium ligands indicated by dotted lines. (c) Proximal calcium site of PNP with the equivalent regions of CCP, APX and LIP superimposed. Colors are the same as in (a). (d) Proximal calcium site of PNP in stereo with calcium ligands indicated by dotted lines. (Figure prepared with SETOR[56].) Structure 1996 4, 311-321DOI: (10.1016/S0969-2126(96)00035-4)

Figure 6 Calcium sites. (a) Distal calcium site of PNP with the equivalent regions of CCP, APX and LIP superimposed. Colors are: CCP red, APX green, LIP purple, and PNP gold. (b) Distal calcium site of PNP in stereo with calcium ligands indicated by dotted lines. (c) Proximal calcium site of PNP with the equivalent regions of CCP, APX and LIP superimposed. Colors are the same as in (a). (d) Proximal calcium site of PNP in stereo with calcium ligands indicated by dotted lines. (Figure prepared with SETOR[56].) Structure 1996 4, 311-321DOI: (10.1016/S0969-2126(96)00035-4)

Figure 6 Calcium sites. (a) Distal calcium site of PNP with the equivalent regions of CCP, APX and LIP superimposed. Colors are: CCP red, APX green, LIP purple, and PNP gold. (b) Distal calcium site of PNP in stereo with calcium ligands indicated by dotted lines. (c) Proximal calcium site of PNP with the equivalent regions of CCP, APX and LIP superimposed. Colors are the same as in (a). (d) Proximal calcium site of PNP in stereo with calcium ligands indicated by dotted lines. (Figure prepared with SETOR[56].) Structure 1996 4, 311-321DOI: (10.1016/S0969-2126(96)00035-4)

Figure 7 Glycosylation sites of PNP. The model appears in thick black lines with the 2Fo–Fc electron-density map in thin gray lines. (a) Asn60 of molecule A. (b) Asn144 of molecule B. (Figure prepared with SETOR [56].) Structure 1996 4, 311-321DOI: (10.1016/S0969-2126(96)00035-4)

Figure 7 Glycosylation sites of PNP. The model appears in thick black lines with the 2Fo–Fc electron-density map in thin gray lines. (a) Asn60 of molecule A. (b) Asn144 of molecule B. (Figure prepared with SETOR [56].) Structure 1996 4, 311-321DOI: (10.1016/S0969-2126(96)00035-4)