Structure of milk lactoperoxidase. A study using circular dichroism and difference absorption spectroscopy.

Circular dichroism spectra of milk lactoperoxidase, its fluoride and cyanide derivatives, and those of ferrous lactoperoxidase and its carbonyl and cyanide compounds, were recorded in the wavelength region 200-670 nm. All derivatives have split ellipticity bands, suggesting that lactoperoxidase has a narrow heme pocket that prevents ligands forming linear iron-ligand bonds. Difference absorption spectroscopy of the enzyme in the far-ultraviolet region supports the previously held view that the fifth ligand of the heme iron is histidine. The secondary structure of lactoperoxidase, calculated from the far-ultraviolet circular dichroism spectrum, contains 65% beta-structure, 23% alpha-helix and 12% unordered structure. Reduction of lactoperoxidase with dithionite gives two forms, indicating that after reduction some compound arising from dithionite binds in the vicinity of the heme iron.