Epitope mapping of horseradish peroxidase (isoenzyme C)
| Autor: | Tn, Ammosova, Iv, Ouporov, Rubtsova MYu, Ov, Ignatenko, Am, Egorov, Ekaterina Kolesanova, Ai, Archakov |
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Models
Molecular Binding Sites Protein Conformation Immune Sera Molecular Sequence Data Enzyme-Linked Immunosorbent Assay Peptide Fragments Protein Structure Secondary Protein Structure Tertiary Isoenzymes Epitopes Amino Acid Sequence Sequence Alignment Epitope Mapping Horseradish Peroxidase Protein Binding |
| Zdroj: | Europe PubMed Central |
| Popis: | Peptide scanning (PEPSCAN) was used to determine linear antigenic determinants of horseradish peroxidase isoenzyme C (HRPC). For this purpose, we synthesized 303 overlapping hexapeptide fragments (with a step of one amino acid residue) of the protein primary structure and studied their interactions with anti-HRPC polyclonal antisera by ELISA. Experiments with various titers of antisera allowed us to determine linear antigenic determinants of HRPC; several such determinants were spatially located in regular elements of the secondary structure (alpha-helices) found both inside and outside the protein globule. A fraction of epitopes were located in loops and folds of the HRPC peptide chain with irregular shapes. These epitopes contained several functionally important residues: Arg 38, which is part of the active site of the enzyme, as well as Phe 142 and Phe 143, which form a channel allowing aromatic substrates to reach the active site. Amino acid residues that form calcium-binding sites or occur in the vicinity of disulfide bonds are not involved in these epitopes. |
| Databáze: | OpenAIRE |
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