Catechol(amine)s as probes of lactoperoxidase catalytic site structure: spectroscopic and modeling studies

• Published in: Journal of biological inorganic chemistry 1999-02, Vol.4 (1), p.12-20
• Main Authors: Ferrari, R P, Traversa, S, De Gioia, L, Fantucci, P, Suriano, G, Ghibaudi, E M
• Format: Article
• Language: English
• Subjects: 3,4-Dihydroxyphenylacetic Acid - chemistry; 3,4-Dihydroxyphenylacetic Acid - metabolism; Animals; Caffeic Acids - chemistry; Caffeic Acids - metabolism; Catalytic Domain; Catecholamines - chemistry; Catecholamines - metabolism; Cattle; Computer Simulation; Dihydroxyphenylalanine - chemistry; Dihydroxyphenylalanine - metabolism; Dopamine - chemistry; Dopamine - metabolism; Hydrogen-Ion Concentration; Hydroxybenzoates - chemistry; Hydroxybenzoates - metabolism; Lactoperoxidase - chemistry; Lactoperoxidase - metabolism; Models, Molecular; Norepinephrine - chemistry; Norepinephrine - metabolism; Protein Conformation; Spectrophotometry, Ultraviolet; Thermodynamics
• ISSN: 0949-8257; 1432-1327
• DOI: 10.1007/s007750050284

Binding affinities to lactoperoxidase (LPO) of a homologous series of substituted catechol(amine)s [such as catechol, 4-methylcatechol, 3,4-dihydroxybenzoic acid, 3,4-dihydroxyphenylacetic acid, 3-(3,4-dihydroxyphenyl)propionic acid; dopamine, noradrenaline, adrenaline; L-3,4-dihydroxyphenylalanine] were studied by UV-visible spectroscopy and docking simulations. Dissociation constant (Kd) values were calculated by direct fitting of the experimental data and fall in a range of 3-95 mM. Thermodynamic parameters are comparable with those reported for the interaction of LPO with p-substituted phenols, suggesting a similar general mode of binding. Furthermore, the relative contributions to binding energy, described by the unimolecular constant Ku, show that interaction between protein and ligands originates from a relatively large number of groups. Docking and molecular dynamics simulations, in agreement with experimental evidence, predict that the substrate is localized into the access channel in the vicinity of heme distal pocket. This channel is characterized by a hydrophobic patch (six Phe residues) and by a charged contribution (two Glu and one His residues). All of the substrates, except caffeic acid, may approach the protein active site. Positively charged Arg372 acts as a gate above the heme distal pocket and seems to address substrate orientation in relation to the side-chain terminal group.