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Proton and iodine-127 nuclear magnetic resonance studies on the binding of iodide by lactoperoxidase

Journal Article · · Biochemistry; (United States)
DOI:https://doi.org/10.1021/bi00394a028· OSTI ID:5599484

Interaction of an iodide ion with lactoperoxidase was studied by the use of /sup 1/H NMR, /sup 127/I NMR, and optical difference spectrum techniques. /sup 1/H NMR spectra demonstrated that a major broad hyperfine-shifted signal at about 60 ppm, which is ascribed to the heme peripheral methyl protons, was shifted toward high field by adding KI, indicating the binding of iodide to the active site of the enzyme; the dissociation constant was estimated to be 38 mM at pH 6.1. The binding was further detected by /sup 127/I NMR, showing no competition with cyanide. Both /sup 1/H NMR and /sup 127/I NMR revealed that the binding of iodide to the enzyme is facilitated by the protonation of an ionizable group with a pK/sup a/ value of 6.0-6.8, which is presumably the distal histidyl residue. Optical difference spectra showed that the binding of an aromatic donor molecule to the enzyme is slightly but distinctly affected by adding KI. On the basis of these results, it was suggested that an iodide ion binds to lactoperoxidase outside the heme crevice but at the position close enough to interact with the distal histidyl residue which possibly mediates electron transport in the iodide oxidation reaction.

Research Organization:
Chiba Univ., Japan
OSTI ID:
5599484
Journal Information:
Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 26:20; ISSN BICHA
Country of Publication:
United States
Language:
English