Horseradish structure

In some cases, the anionic POs adsorbed on chitin have similar antigenic determinants, but the plants belonging to different families - and even members of the same family - could have polysaccharide-specific POs with different structures. Thus, the majority of investigated species had anionic chitin-specific peroxidises, and these isoforms from potato Solcmaceae) and horseradish Brassicaceae) formed lines of precipitation with antibodies to wheat chitin-bound PO but not to anionic isoPO (Maksimov et al., 2000). However, protein extracts from several plants of Brassicaceae, Cucurhitaceae and Fahaceae formed precipitate with both the chitin-specific and anionic PO of wheat (Fig. 3). It was foimd that the greatest homology showed in plants and formed precipitation lines with the anionic PO of wheat (Tab. 2). 

As a result of the micellar environment, enzymes and proteins acquire novel conformational and/or dynamic properties, which has led to an interesting research perspective from both the biophysical and the biotechnological points of view [173-175], From the comparison of some properties of catalase and horseradish peroxidase solubilized in wa-ter/AOT/n-heptane microemulsions with those in an aqueous solution of AOT it was ascertained that the secondary structure of catalase significantly changes in the presence of an aqueous micellar solution of AOT, whereas in AOT/n-heptane reverse micelles it does not change. On the other hand, AOT has no effect on horseradish peroxidase in aqueous solution, whereas slight changes in the secondary structure of horseradish peroxidase in AOT/n-heptane reverse micelles occur [176],... 

Due to the relatively high-molecular-weight of the enzyme, conjugates formed with antibodies and P-gal tend to be much bulkier than those associated with AP or horseradish peroxidase. For this reason, antibody conjugates made with P-gal may have more difficulty penetrating tissue structures during immunohistochemical staining techniques than those made with the other enzymes. 

Dunford HB. Horseradish peroxidase structure and kinetic properties. In Everse I, Everse KE, Grisham MB, eds. Peroxidases in Chemistry and Biology. Boca Raton, FL CRC Press 1991 1-24. 

APX, ascorbate peroxidase PJiP, Arthromyces ramosus peroxidase BPl, barley grain peroxidase CCP, C3dochrome c peroxidase CIP, Coprinus cinereus peroxidase EXAFS, extended X-ray absorption fine structure HRP, horseradish peroxidase HRP Z (where Z = A1-A3, B1-B3, Cl, C2, D, E1-E6, or N), a specific isoenzyme of horseradish peroxidase HS, high-spin lAA, indole-3-acetic acid LIP, hgnin peroxidase LS, low-spin PNP, the major cationic isoenzyme of peanut peroxidase WT, wild-type 5-c, five-coordinate 6-c, six-coordinate. 

Fig. 2. Nomenclature of the heme group of horseradish peroxidase according to the lUB/IUPAC system. Proton-bearing carbon atoms are numbered on the structure and can be cross-referenced with the Fischer system as follows ...

The presence of calcium in horseradish peroxidase was demonstrated originally by Haschke and Friedhoff, working with the C and A (imspec-ified, but likely to have been predominantly A2) isoenzymes (209). HRP C and HRP A contain 2.0 0.13 and 1.4 0.19 moles calcium per mole enzyme, respectively, as determined by atomic absorption spectroscopy. Incubation in 6 M guanidinium hydrochloride and 10 mM EDTA for 4 hours at neutral pH and room temperature gave calcium-depleted enzymes with specific activities decreased by 40% and 15%, respectively. The thermal stability of calcium-depleted HRP C was also reduced compared to native enzyme. Reconstitution was successful only with calcium-depleted HRP C (209). It remains to be established whether this reflects true structural differences between the calcium binding sites of the two isoenz5unes, or is a consequence of the relatively harsh... 

---