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Electron paramagnetic resonance spectroscopy of lactoperoxidase complexes: clarification of hyperfine splitting for the NO adduct of lactoperoxidase

Journal Article · · Biochemistry; (United States)
OSTI ID:5589154

Electron paramagnetic resonance (EPR) studies of the nitrosyl adduct of ferrous lactoperoxidase (LPO) confirm that the fifth axial ligand in LPO is bound to the iron via a nitrogen atom. Complete reduction of the ferric LPO sample is required in order to observe the nine-line hyperfine splitting in the ferrous LPO/NO EPR spectrum. The ferrous LPO/NO complex does not exhibit a pH or buffer system dependence when examined by EPR. Interconversion of the ferrous LPO/NO complex and the ferric LPO/NO/sub 2//sup -/ complex is achieved by addition of the appropriate oxidizing or reducing agent. Characterization of the low-spin LPO/NO/sub 2//sup -/ complex by EPR and visible spectroscopy is reported. The pH dependence of the EPR spectra of ferric LPO and ferric LPO/CN/sup -/ suggests that a high-spin anisotrophic LPO complex is formed at high pH and an acid-alkaline transition of the protein conformation near the heme site does occur in LPO/CN/sup -/. The effect of tris(hydroxymethyl)aminomethane buffer on the LPO EPR spectrum is also examined.

Research Organization:
Univ. of Iowa, Iowa City
OSTI ID:
5589154
Journal Information:
Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 26:22; ISSN BICHA
Country of Publication:
United States
Language:
English