Реферат: In order to reveal the conformational changes in holo- and apo-forms of horseradish peroxidase (HRP) the di erences in immunogenicity of these forms have been studied by immunochemical techniques. A panel of polyclonal antibodies (PcAb) speci c to holo- and apo-HRP was tested by double immunodi usion (Ouchterlony test) to nd the antisera which distinguish the structural changes in peroxidase molecule after heme removal. Two sera speci c to apo-HRP were revealed to recognize the epitopes speci c only for apo-HRP. Peptide scanning (PEPSCAN) of these PcAb was used to determine linear antigenic determinants for apo-form and compare them with those obtained for holo-HRP earlier. We found most epitopes established for holo-HRP, but some of them were shifted by one-two amino acids. At the same time we revealed several new epitopes speci c only for apo-HRP. Some of epitopes placed close to the active site were speci c only for holo-HRP. The increased amount of antigenic epitopes established for apo-HRP and partial shifts in their location mean certain conformational changes in peroxidase molecule in a result of heme removal. It also means that apo-HRP is less compact protein with increased exibility of segments and loops. These facts con rm the role of heme in close approach of two domains and stabilization of enzyme molecule.

Библиографическая ссылка: Ignatenko O.V. , Rubtsova M.Y. , Ivanova N.L. , Ouporov I.V. , Egorov A.M.
Analysis of Antigenic Structure of Holo- and Apo-forms of Horseradish Peroxidase
Вестник Московского Университета. Серия 2: Химия. 2000. V.41. N6 (Supplement). P.102-106.

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