It is possible now to follow heme-histidine contacts by characteristic absorption bands in second derivative spectra in Soret region. The method gives the possibility to find out heme-tryptophan contact in UV region in some proteins e.g. horseradish peroxidase. The differences revealed in the positions of the absoption bands in second derivative spectra can be helpful for the definition of the peculiarities of heme liganding in different heme containing proteins. Second derivative spectra allows to discriminate individual absorption components of chromophoric groups in peroxidase, e.g. aromatic amino acid residues and coordinated heme. The correlation between spectral characteristics and coordination state and charge of heme in peroxidase was suggested using of heme-containing model systems.
First stage is the process of heme modification.
Modified enzyme possesses peroxidase activity. Second stage consists of destruction of heme derivative and apo-enzyme formation. The obtained monoclonal antibodies were IgG1 and did not inhibit the enzyme activity at the concentrations precluding the precipitate formation. Seven monoclonal antibodies interacted with basic isozymes, while two of them - with acidic isozymes. Monoclonal and polyclonal antibodies to acidic isoforms of peroxidase did not interact with basic isozymes, and to basic isoforms with acidic ones.