title: Kinetic evidence supports the existence of two halide binding sites that have a distinct impact on the heme iron microenvironment in myeloperoxidase.
abstract: Myeloperoxidase (MPO) structural analysis has suggested that halides and pseudohalides bind to the distal binding site and serve as substrates or inhibitors, while others have concluded that there are two separate sites. Here, evidence for two distinct binding sites for halides comes from the bell-shaped effects observed when the second-order rate constant of nitric oxide (NO) binding to MPO was plotted versus Cl- concentration. The chloride level used in the X-ray structure that produced Cl- binding to the amino terminus of the helix halide binding site was insufficient to populate either of the two sites that appear to be responsible for the two phases. Biphasic effects were also observed when the I-, Br-, and SCN- concentrations were plotted against the NO combination rate constants. Interestingly, the trough concentrations obtained from the bell-shaped curves are comparable to normal plasma levels of halides and pseudohalides, suggesting the potential relevance of these molecules in modulating MPO function. The second-order rate constant of NO binding in the presence of plasma levels of I-, Br-, and SCN- is 1-2-fold lower compared to that obtained in the absence of these molecules and remains unaltered through the Cl- plasma level. When Cl- exceeded the plasma level, the NO combination rate becomes indistinguishable from the second phase of the bell-shaped curve that was obtained in the absence of halides. Our results are consistent with two halide binding sites that could be populated by two halides in which both display distinct effects on the MPO heme iron microenvironment.
authors.affiliation: Department of Obstetrics and Gynecology, The C. S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
authors.forename: Gheorghe
authors.initials: G
authors.lastname: Proteasa
authors.forename: Yahya R
authors.initials: YR
authors.lastname: Tahboub
authors.forename: Semira
authors.initials: S
authors.lastname: Galijasevic
authors.forename: Frank M
authors.initials: FM
authors.lastname: Raushel
authors.forename: Husam M
authors.initials: HM
authors.lastname: Abu-Soud
mesh_terms: D000838:Anions / Q000378:metabolism; D001665:Binding Sites; D002712:Chlorides / Q000378:metabolism; D004795:Enzyme Stability; D006418:Heme / Q000737:chemistry; D006801:Humans; D066298:In Vitro Techniques; D007501:Iron / Q000737:chemistry; D007700:Kinetics; D008958:Models, Molecular; D009569:Nitric Oxide / Q000378:metabolism; D009195:Peroxidase / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D000838:Anions; D002712:Chlorides; D009569:Nitric Oxide; D006418:Heme; D007501:Iron; D009195:Peroxidase