The first evidence of multi-component complexes formed by myeloperoxidase (MPO), ceruloplasmin (CP), and very low/low density lipoproteins (VLDL/LDL) obtained by electrophoresis, gel filtration, and photon-correlation spectroscopy (PCS) is...
The first evidence of multi-component complexes formed by myeloperoxidase (MPO), ceruloplasmin (CP), and very low/low density lipoproteins (VLDL/LDL) obtained by electrophoresis, gel filtration, and photon-correlation spectroscopy (PCS) is presented in this paper. Complexes were observed when isolated MPO, CP, and VLDL/LDL were mixed and/or when MPO was added to the blood plasma. Complex LDL-MPO-CP was detected in 44 of 100 plasma samples taken from patients with atherosclerosis, and 33 of 44 samples also contained the VLDL-MPO-CP complex. MPO concentration in these patients' plasma exceeded 800 ng/ml. Interaction of MPO with high density lipoproteins (HDL) was not revealed, as well as binding of CP to lipoproteins in the absence of MPO. Adding antibodies against apoB-100 to VLDL-MPO-CP and LDL-MPO-CP complexes results in release of lipoproteins. Using PCS the diameters of complexes under study were evaluated. By comparing concentrations of the components in complexes formed by MPO, CP, and lipoproteins their stoichiometry was assessed as 2VLDL:1MPO:2CP and 1LDL:1MPO:2CP. Lipoproteins affected the inhibition of MPO peroxidase activity by CP. The affinity of lipoproteins to MPO-CP complex was assessed using apparent dissociation constants determined as approximately 0.3 nM for VLDL and approximately 0.14 nM for LDL. Sokolov A.V., Ageeva K.V., Cherkalina O.S., Pulina M.O., Zakharova E.T., Prozorovskii V.N., Aksenov D.V., Vasilyev V.B., Panasenko O.M. (2010) Identification and properties of complexes formed by myeloperoxidase with lipoproteins and ceruloplasmin. Chem. Phys. Lipids. V. 163. P. 347-355.
Sokolov AV, Ageeva KV, Pulina MO, Cherkalina OS, Samygina VR, Vlasova II, Panasenko OM, Zakharova ET, Vasilyev VB.
- Free Radical Research , 2008
Ceruloplasmin (CP), the multicopper oxidase of plasma, interacts with myeloperoxidase (MPO), an enzyme of leukocytes, and inhibits its peroxidase and chlorinating activity. Studies on the enzymatic properties shows that CP behaves as a competitive...
Ceruloplasmin (CP), the multicopper oxidase of plasma, interacts with myeloperoxidase (MPO), an enzyme of leukocytes, and inhibits its peroxidase and chlorinating activity. Studies on the enzymatic properties shows that CP behaves as a competitive inhibitor impeding the binding of aromatic substrates to the active centre of MPO. The contact between CP and MPO probably entails conformational changes close to the p-phenylenediamine binding site in CP, which explains the observed activation by MPO of the substrate's oxidation. CP subjected to partial proteolysis was virtually unable to inhibit activity of MPO. The possible protein-protein interface is comprised of the area near active site of MPO and the loop linking domains 5 and 6 in CP. One of the outcomes of this study is the finding of a new link between antioxidant properties of CP and its susceptibility to proteolysis. Sokolov, A.V., Ageeva, K.V., Pulina, M.O., Cherkalina, O.S., Samygina, V.R., Vlasova, I.I., Panasenko, O.M., Zakharova, E.T., Vasilyev, V.B. (2008) Ceruloplasmin and myeloperoxidase in complex affect the enzymatic properties of each other.// Free Radic. Res. V. 42. P. 989-998
Sokolov AV, Ageeva KV, Pulina MO, Zakharova ET, Vasilyev VB.
- Biometals , 2009
In our previous report we first described a complex between lactoferrin (Lf) and ceruloplasmin (Cp) with K (d) ~ 1.8 muM. The presence of this complex in colostrum that never contains more than 0.3 muM Cp questions the reliability of K (d) value....
In our previous report we first described a complex between lactoferrin (Lf) and ceruloplasmin (Cp) with K (d) ~ 1.8 muM. The presence of this complex in colostrum that never contains more than 0.3 muM Cp questions the reliability of K (d) value. We carefully studied Lf binding to Cp and investigated the enzymatic activity of the latter in the presence of Lf, which allowed obtaining a new value for K (d) of Cp-Lf complex. Lf interacting with Cp changes its oxidizing activity with various substrates, such as Fe(2+), o-dianisidine (o-DA), p-phenylenediamine (p-PD) and dihydroxyphenylalanine (DOPA). The presence of at least two binding sites for Lf in Cp molecule is deduced from comparison of substrates' oxidation kinetics with and without Lf. When Lf binds to the first site affinity of Cp to Fe(2+) and to o-DA increases, but it decreases towards DOPA and remains unchanged towards p-PD. Oxidation rate of Fe(2+) grows, while that of o-DA, p-PD and DOPA goes down. Subsequent Lf binding to the second center has no effect on iron oxidation, hampers DOPA and o-DA oxidation, and reduces affinity towards p-PD. Scatchard plot for Lf sorbing to Cp-Sepharose allowed estimating K (d) for Lf binding to high-affinity (~13.4 nM) and low-affinity (~211 nM) sites. The observed effect of Lf on ferroxidase activity of Cp is likely to have physiological implications. Sokolov AV, Ageeva KV, Pulina MO, Zakharova ET, Vasilyev VB. (2009) Effect of lactoferrin on oxidative features of ceruloplasmin.// Biometals, V. 22, P. 521-529.
