Similarities and differences of five peroxidases from turnip and horseradish. Peptide mapping studies on glycoproteins.
Four isoperoxidases of turnip root and isoperoxidase C of horseradish root were digested with trypsin, and their peptide maps, prepared by high-voltage paper electrophoresis, were compared. All five tryptic digests were completely soluble at pH 8. The maps were developed with a variety of general and specific reagents: ninhydrin, histidine, tyrosine, tryptophan and arginine reagents. Cystine peptides and cysteic acid derivatives have also been characterized. All detected half-cystine residues seemed engaged in disulfide bridges. For each individual peroxidase the number of specifically staining peptides agreed very well with the amino acid composition. The two most acidic peroxidases of turnip, P1 and P2, only differ significantly in one peptide. The P2 gene is tentatively proposed to have developed from the P1 gene by a single base mutation, changing an asparagine residue to alysine residue. A less acidic turnip peroxidase, P3, is distinct, although related to peroxidases P1 and P2. Horseradish isoperoxidase C also belongs to this group which appears to be closely related in the amino acid sequences around four disulfide bridges. Peroxidase P7 differs from this group, at least around two of its disulfide bridges, and therefore, may differ from the other four in parts of its three dimensional structure. Sequences of particular importance to peroxidase function must be present in all peroxidases. From the peptide mapping studies we only find two highly homologous sequences present in all five examined peroxidases. Both contain histidine. This finding corroborates previous suggestions of two histidine sequences near the peroxidase heme prosthetic group. The rules applied in relating peptides of different proteins are outlined, and the sources of errors in mapping of glycoproteins of high carbohydrate content (about 20%) are discussed in detail.
D000596:Amino Acids / Q000032:analysis; D005944:Glucosamine / Q000032:analysis; D006023:Glycoproteins / Q000032:analysis*; D006735:Horseradish Peroxidase / Q000032:analysis*; D007527:Isoenzymes / Q000032:analysis*; D010446:Peptide Fragments / Q000032:analysis; D010544:Peroxidases / Q000032:analysis*; D010944:Plants / Q000032:analysis*; D013045:Species Specificity; D014357:Trypsin
D000596:Amino Acids; D006023:Glycoproteins; D007527:Isoenzymes; D010446:Peptide Fragments; D006735:Horseradish Peroxidase; D010544:Peroxidases; D014357:Trypsin; D005944:Glucosamine