Influence of hemoglobin and albumin on the NO donation effect of tetranitrosyl iron complex with thiosulfate
Introduction
Structural synthetic analogs of the active center of non-heme iron-sulfur proteins, nitrosyl iron complexes (DNICs) with functional sulfur-containing ligands, are of great interest as new generation drugs for the treatment of cardiovascular, oncological and other socially significant diseases [1,2].
Anionic binuclear DNIC with thiosulfate ligands (complex 1) Na2 [Fe2(S2O3)2(NO)4]∙4H2O (Fig. 1) is one of the most promising representatives of this compounds class [3]. It has been shown that complex 1 is a more effective NO donor compared with the NONOate, 2,2′-(hydroxynitrosohydrazono)bis-ethanimine [4], it exhibits antimetastatic activity and induces the expression of the genes of the protective DNA repair pathways in the cell, including the SOS and SoxRS systems of E. Coli [5], and also completely inhibits the catalytic activity of Ca2+-ATPase and phosphodiesterase at a concentration of 0.01 mM [6].
To understand the chemical and biological mechanisms of complex 1 action as a prodrug, a comprehensive study of its metabolic processes is necessary to conduct. It is assumed that the targets of DNICs as NO donors in vivo can be Fe- and thiol-containing proteins, as well as low molecular weight thiols [7,8]. In this work, the biotransformation of complex 1 was analyzed by the effects of its interaction with deoxyhemoglobin (Hb) and bovine serum albumin (BSA).
According to the literature [9], erythrocyte hemoglobin is one of the most important elements of the biological transformation of NO donors and the NO transport inside the body. It was found that besides the formation of protein-bound complexes with the oxygenated form of the protein (HbO2) [10], DNICs are also able to bind with deoxygenated Hb [11], becoming more prolonged NO donors. As a result of the analysis carried out in Ref. [11], it was concluded that the observed effect of cationic complexes stabilization can occur due to the binding of their positively charged groups with negatively charged amino acid residues on the Hb surface. It was interesting to determine whether a similar effect would be observed in the case of anionic complex 1.
Another no less important objective of this work was to study the effect of BSA on the decay process of complex 1. BSA is a blood transport protein the main function of which is a transfer of various biologically active substances [12]. It has been established [7] that BSA is one of the main targets of the DNICs action, and the binding of DNICs occurs with cysteine (Cys 34) or histidine residues in the hydrophobic pocket of the molecule to form protein-bound nitrosyl complexes [13].
According to experimental data obtained in animals [14], high-molecular complexes formed by the interaction of DNICs with various proteins are more stable than low-molecular complexes, and as a result, most of the DNICs in cells and tissues have a protein form.
Section snippets
Materials
In the work, tris(hydroxymethyl)aminomethane (“Serva”, Germany), sulfanilamide (SA) (“Sigma”, USA), N-(1-Naphthyl)ethylenediamine dihydrochloride (NEDA) (“MP Biomedicals”, Germany) and bovine serum albumin, fraction 5 (BioClot, GmbH) were used. Water was distilled in a “Bi/Duplex” distiller (Germany). Binuclear tetranitrosyl iron complex 1 has been obtained using techniques [3]. A homogeneous solution of HbO2 was isolated from fresh blood of experimental animals by the known method [15]. To
Results
Hb is very convenient for the spectrophotometric recording of the NO generation kinetics from complex 1 because it is a trap for NO: the rate of NO binding to the Hb heme is close to diffusion, the equilibrium constant is 3∙1010 M−1 [17]. In addition, formed nitrosyl hemoglobin (HbNO) has a characteristic absorption spectrum: one Hb absorption maxima (556 nm) splits into two (545 nm and 575 nm) during HbNO formation, which is convenient to use to determine the depth of the reaction. Thus, the
Discussion
The results of this work demonstrate a high stabilizing ability of proteins towards the complex 1. As was shown earlier, the albumin-bound complex is formed by the rapid reaction of free cysteine and histidine groups with DNIC [13]. Hemoglobin, from this point of view, is a unique protein that can bind DNICs and their decay products in various ways [10,23]. In arterial blood, where HbO2 predominates, the binding of NO and DNICs occurs with the free SH group (Cys 93). At the same time, when the
Acknowledgements
The work was supported by the Ministry of Education and Science of the Russian Federation, state task No 0089-2019-0014 and the Presidium Program of the Russian Academy of Sciences for 2018–2020. “Nanostructures: physics, chemistry, biology, fundamentals of technology”, the project “Development of ways to optimize the pharmacological properties of biologically active compounds through inclusion in molecular nanostructures”.
References (24)
- et al.
Dinitrosyl iron complexes bind with hemoglobin as markers of oxidative stress
- et al.
Transformation of mononuclear dinitrosyl iron complex (DNIC) with thiourea in glutathione aqueous solution
J. Mol. Struct.
(2019) - et al.
S-nitrosation of serum albumin by dinitrosyl-iron complex
J. Biol. Chem. V.
(1995) - et al.
Preparation and properties of α- and β-chains from human hemoglobin
J. Biol. Chem.
(1969) - et al.
Reactions of sulfur-nitrosyl iron complexes of “g=2.03” family with hemoglobin (Hb): kinetics of Hb–NO formation in aqueous solutions
Nitric Oxide
(2007) - et al.
Determination of nitric oxide using fluorescence spectroscopy
Methods Enzymol.
(1996) - et al.
Polynuclear water-soluble dinitrosyl iron complexes with cysteine or glutathione ligands: electron paramagnetic resonance and optical studies
Nitric Oxide
(2010) - et al.
A new class of nitric oxide donors
Her. Russ. Acad. Sci.
(2016) - et al.
Nitrosyl iron complexes—synthesis, structure and biology
Dalton Trans.
(2011) - et al.
Synthesis, structure and solid-phase transformations of Fe nitrosyl complex Na2[Fe2(S2O3)2(NO)4]·4H2O
Russ. J. Coord. Chem.
(2005)
Antitumor activity of iron nitrosyl complexes: new donors of nitrogen monoxide
Ross. Kh. Zh.
formation of dinitrosyl iron complex is a necessary step in the realization of the Na2[Fe2(S2O3)2(NO)4]·4H2O genetic activity
Dokl. Biol. Sci.
Cited by (7)
Anionic dinitrosyl iron complexes – new nitric oxide donors with selective toxicity to human glioblastoma cells
2022, Journal of Molecular StructureCitation Excerpt :The kinetic curves of NO release reach a plateau for both complexes by ∼300 second of the experiment, then the systems reach equilibrium both in the case of buffer (pH 7.0) solutions (Fig. 5, curves 1a, 1b) and in the case of DMEM medium (Fig. 5, curves 2a, 2b). The observed decrease in NO generation rates in DMEM compared to that observed in PBS could be explained by the presence of serum in the medium, since serum proteins stabilize DNICs and prolong NO release [73–75]. Influence of other components of DMEM on the NO generation can not be excluded as well.
Effects of albumin-bound nitrosyl iron complex with thiosulfate ligands on lipid peroxidation and activities of mitochondrial enzymes in vitro
2021, Nitric Oxide - Biology and ChemistryCitation Excerpt :Plots, linear regressions were determined using Origin 6.1 for Windows, OriginLab (Northampton, MA, USA). It was previously found [18,19] that bovine serum albumin can stabilize TNIC effectively, which makes it decay 27 times slower and leads to its more prolonged action as a NO donor. The studies of biological effects, to which this work is devoted, are carried out in the air; therefore, the stability of TNIC in the system with albumin under these conditions was assessed.
Features of the decomposition of cationic nitrosyl iron complexes with N-ethylthiourea and penicillamine ligands in the presence of albumin
2021, Inorganica Chimica ActaCitation Excerpt :We have previously studied the effect of bovine serum albumin (BSA) on the decomposition of an anionic binuclear complex with thiosulfate ligands [35] and a cationic mononuclear complex with thiourea ligands in vitro [36]. It was found in [35] that the complex decomposes in the presence of BSA 27 times slower (under anaerobic conditions). In [36], the binding sites of the complex on the protein surface were analyzed using the molecular docking method.
Nitrosyl iron complex with N-ethylthiourea ligands: reactions with hemoglobin
2023, Russian Chemical Bulletin