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Biomedical subjects

M Soszyński

Publications and source records attributed to M Soszyński.

At least 19 recordsLinked to original sources
PubMed2000

Dopamine-melanin protects against tyrosine nitration, tryptophan oxidation and Ca(2+)-ATPase inactivation induced by peroxynitrite.

The effects of dopamine-melanin (DA-melanin), a synthetic model of neuromelanin, on peroxynitrite-mediated 3-nitrotyrosine formation, oxidation of tryptophan in bovine serum albumin and inactivation of erythrocyte membrane Ca(2+)-ATPase activity were investigated in the absence and in the presence of bicarbonate. DA-melanin inhibited nitration of free tyrosine, loss of tryptophan residues and Ca(2+)-ATPase inactivation by peroxynitrite in a dose dependent manner. In the presence of bicarbonate, this inhibitory effect was lower for nitration and insignificant for oxidative protein modifications. These results suggest that neuromelanin can protect against nitrating and oxidizing action of peroxynitrite but is a worse protector against the peroxynitrite-CO(2) adduct. As peroxynitrite may be a mediator of neurotoxic processes, the obtained results suggest that neuromelanin may be important as a physiological protector against peroxynitrite.

Animals↗
PubMed2000

Inactivation of antioxidant enzymes by peroxynitrite.

Exposure of hemolysates and whole erythrocytes to peroxynitrite (bolus of 50 micromol dm(-3)-2 mmol dm(-3)) was found to inactivate erythrocyte antioxidant enzymes: glutathione peroxidase > superoxide dismutase > catalase. Inactivation of antioxidant enzymes by peroxynitrite may be one reason for the secondary oxidative stress in peroxynitrite-treated cells. When hemoglobin was not converted into the cyanmet form, an apparent activation of glutathione peroxidase activity by peroxynitrite was observed in hemolysates; this effect was artifactual and due to the pseudoenzymatic glutathione peroxidase activity of hemoglobin.

Catalase↗
PubMed2000

Plasma membrane Ca2+-ATPase in excitable and nonexcitable cells.

There is a significant number of data confirming that the maintenance of calcium homeostasis in a living cell is a complex, multiregulated process. Calcium efflux from excitable cells (i.e., neurons) occurs through two main systems--an electrochemically driven Na+/Ca2+ exchanger with a low Ca2+ affinity (K0.5 = 10-15 microM), and a plasmalemmal, specific Ca2+-ATPase, with a high Ca2+ affinity (K0.5 < 0.5-1 microM), whereas in nonexcitable cells (i.e., erythrocytes) the calcium pump is the sole system responsible for the extrusion of calcium ions. The plasma membrane Ca2+-ATPase (PMCA) is a ubiquitously expressed protein, and more than 26 transcripts of four PMCA genes are distributed in a tissue specific manner. Differences in the structure and localization of PMCA variants are thought to correlate with specific regulatory properties and may have consequences for proper cellular Ca2+ signaling. The regulatory mechanisms of calcium pump activity have been studied extensively, resulting in a new view of the functioning of this important molecule in the membranes.

Animals↗
PubMed2000

Transport of organic anions by multidrug resistance-associated protein in the erythrocyte.

The active transport of oxidized glutathione and glutathione S-conjugates has been demonstrated for the first time in erythrocytes and this cell remained the main subject of research on the "glutathione S-conjugate pump" for years. Further studies identifled the "glutathione S-conjugate pump" as multidrug resistance-associated protein (MRP). Even though cells overexpressing MRP and isolated MRP provide useful information on MRP structure and function, the erythrocyte remains an interesting model cell for studies of MRP1 in its natural environment, including the substrate specificity and ATPase activity of the protein.

ATP-Binding Cassette Transporters↗
PubMed1998

Radiation inactivation suggests that human multidrug resistance-associated protein 1 occurs as a dimer in the human erythrocyte membrane.

Molecular masses of functional units of two components of 2, 4-dinitrophenyl-S-glutathione (DNP-SG) transport across the erythrocyte membrane determined by radiation inactivation were 437 +/- 69 kDa for the high-affinity component and 466 +/- 67 kDa for the low-affinity component. These results confirm that the multidrug resistance-associated protein (MRP) 1 is responsible for the high-affinity DNP-SG transport across the erythrocyte membrane and suggest that MRP1 exists in the membrane as a dimer. The molecular size of the low-affinity transporter is similar if not identical to that of MRP1. Moreover, while the molecular mass of the DNP-SG-ATPase activity of the erythrocyte membrane corresponds also to that of MRP (375 +/- 36 kDa), the molecular mass of the functional unit of dinitrophenol-stimulated ATPase is significantly lower (232 +/- 26 kDa), which suggests that thisactivity is linked to a different protein, perhapsaminophospholipid translocase.

2,4-Dinitrophenol↗
PubMed1998

Effect of ethanol and formate radicals on erythrocyte membrane proteins.

PURPOSE: The effect of ethanol and formate radicals on the major proteins of human erythrocyte membranes has been investigated. MATERIALS AND METHODS: Human erythrocyte ghosts and of erythrocyte ghosts stripped of peripheric proteins were irradiated in phosphate buffer with 100 mmol dm(-3) ethanol or 100 mmol dm(-3) formate under N2 or N2O. The alterations of the proteins were investigated by SDS-polyacrylamide gel electrophoresis and high-performance gel permeation chromatography. RESULTS: In contrast to previous results on ribonuclease and on serum albumin the ethanol radicals were found to have a higher efficiency to damage erythrocyte membrane proteins than the formate radicals. Spectrin (Bands 1 and 2) and capnophorin (Band 3) showed the highest radiation-induced loss of all membrane proteins. When cysteamine or dithiothreitol were added to the erythrocyte ghosts with a similar OH-scavenging capacity as ethanol or formate, no degradation or aggregation of the membrane proteins could be observed even after a dose as high as 1800 Gy. CONCLUSIONS: The results of this study confirm the high radiosensitivity of spectrin and capnophorin to primary radicals. Similarly to soluble proteins, membrane-associated proteins are more significantly damaged by ethanol radicals than by formate radicals.

Adult↗
PubMed1997

Peroxynitrite inhibits glutathione S-conjugate transport.

Peroxynitrite was demonstrated to inhibit the active efflux of glutathione S-conjugates (2,4-dinitrophenyl-S-glutathione and bimane-S-glutathione) from human erythrocytes and the erythrocyte membrane ATPase activity stimulated by glutathione S-conjugates. As the multidrug resistance-associated protein (MRP) is responsible for the transport of glutathione S-conjugates in mammalian cells, these results point to the possibility of the effect of peroxynitrite on the MRP function.

Biological Transport↗
PubMed1997

Decrease in accessible thiols as an index of oxidative damage to membrane proteins.

The effect of several oxidative agents (hydrogen peroxide, tert-butyl hydroperoxide, menadione, AAPH, peroxynitrite and ionizing radiation) on the ratio of weakly to strongly immobilized residues of erythrocyte membrane-bound maleimide-tempo spin label (h(w)/h(s) ratio) was studied in order to test the hypothesis that a decrease in the h(w)/h(s) ratio may be a general index of oxidative damage to membrane proteins. Most of the agents studied decreased though H2O2 almost did not affect and ionizing radiation increased the h(w)/h(s) ratio. In parallel, the ratio of DTNB accessible/DTNB inaccessible membrane protein-SH groups was determined from membrane-SH group measurements with the Ellman reagent in the absence and in the presence of sodium dodecyl sulfate. This ratio decreased in all cases studied and seems to be a more universal and easy to measure parameter to describe the oxidative damage to membrane proteins.

Adult↗
PubMed1997

Effect of postirradiation treatment on the radiation-induced haemolysis of human erythrocytes.

The effect of postirradiation conditions on the haemolysis of y-irradiated (2.1 kGy) human erythrocytes was studied. Haemolysis was inhibited by incubation in mannitol and sucrose instead of saline, by hypertonicity of the medium and by calcium chelators. Dithiothreitol, butylated hydroxytoluene, deferoxamine, DIDS (in inhibitor of anion exchange) and furosemide (an inhibitor of K/Cl and K/Na/Cl cotransport) did not slow down the haemolysis. Apparently, the radiation-induced haemolysis is due to the formation of membrane pores leaky for electrolytes. From the inhibition of haemolysis by mannitol, apparent pore radius was estimated to be about 0.7 nm. The pores appear to be transient, the average pore number per cell being much less than unity.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1997

Penetration of erythrocyte membrane by peroxynitrite: participation of the anion exchange protein.

Participation of the anion exchange protein (Band 3 protein) of the erythrocyte membrane in the transport of peroxynitrite into erythrocytes was shown by partial inhibition of hemoglobin oxidation by extracellular peroxynitrite in cells treated with Band 3 inhibitors. These results demonstrate that permeation in the anionic form may be a minor pathway in the membrane transport of the peroxynitrite anion/peroxynitrous acid couple, especially in membranes reach in anion exchangers or channels.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1996

Effect of peroxynitrite on erythrocytes.

The action of peroxynitrite on human erythrocytes and erythrocyte membranes was studied. Peroxynitrite (0.1-2 mM) induced a transient decrease of intracellular reduced glutathione, oxidized membrane protein -SH groups, initiated membrane lipid peroxidation and inactivated erythrocyte membrane acetylcholinesterase and ATPase activities. Membranes exposed to peroxynitrite showed aggregation and nitration of proteins and changes in protein organization detectable with a maleimide spin label.

Acetylcholinesterase↗
PubMed1995

Peroxides inhibit the glutathione S-conjugate pump.

Tert-Butyl hydroperoxide (100-300 microM) was found to inhibit the active efflux of dinitrophenyl-S-glutathione (DNP-SG) from human erythrocytes. From among amino acid peroxides generated by irradiation of amino acid (proline, valine and leucine) solutions, valine hydroperoxide (150 microM) had a similar effect on the DNP-SG transport. As the transport of glutathione S-conjugates is an important step of cellular detoxication, these results indicate that oxidative stress may impair cellular resistance to chemical stress of other kinds. t-Butyl and amino acid peroxides upon interaction with hemoglobin and whole erythrocytes produce free radicals which may be responsible for the damage to the glutathione S-conjugate pump of the erythrocyte membrane.

Biological Transport, Active↗
PubMed1991

Effect of X-irradiation on erythrocyte membrane proteins. Primary radicals.

The effect of X-irradiation on the major proteins of human erythrocyte membranes have been examined. Samples of human erythrocyte ghosts and stripped ghosts were irradiated (up to 1.5 kGy) under air, N2 or N2O. The effects on the main erythrocyte membrane proteins as well as on aggregate formation were investigated using sodium dodecyl sulphate-polyacrylamide gel electrophoresis and high-performance gel permeation chromatography. Experiments were carried out with or without dithiothreitol as a reducing agent. The main peripheral protein of the erythrocyte membrane, spectrin, is more radio-sensitive than the other membrane proteins. Degradation was mainly due to aggregation and was increased by excluding oxygen. Since radiolysis under N2O instead of N2 enhanced the loss of spectrin, OH radicals seemed to be especially effective. Under anaerobic conditions the degradation of band 3 material could also be observed. In stripped erythrocyte ghosts the radiosensitivity of this integral protein was similar to that of spectrin.

Air↗
PubMed1988

Self-digestion of erythrocyte membranes of various mammalian species.

1. The rate of self-digestion of main membrane proteins, spectrin and band 3 protein, was studied for erythrocyte membranes of eight mammalian species: man, cow, pig, cat, rabbit, hamster, mouse and rat. 2. Spectrin and band 3 protein were most rapidly degraded in human and pig ghosts. The rates of proteolysis for other species were similar. 3. The rates of self-digestion were correlated neither with lifetime of red cells nor lifespan of animals of various species.

Animals↗
PubMed1987

Aged erythrocytes exhibit decreased anion exchange.

The rate of transport of [32P] phosphate into human and bovine erythrocytes and of a spin-label analogue of phosphate (Tempo-phosphate) into human erythrocytes was found to decrease with increasing erythrocyte age by 15-20% when comparing 20% most dense cells with 20% of lightest cells. The activation energy of Tempo-phosphate transport did not show significant changes upon erythrocyte aging.

Animals↗
PubMed1986

Proteolytic susceptibility of membrane proteins during erythrocyte aging.

Susceptibility of membrane proteins to digestion with chymotrypsin was compared in various age fractions of bovine erythrocytes by SDS-polyacrylamide gel electrophoresis. The extent of protein digestion was found to be higher in membranes of older erythrocytes, as judged from the disappearance of original protein bands and accumulation of digestion products. These results are consistent with the hypothesis of a proteolytic generation of a "senescent cell antigen" in red blood cells.

Animals↗
PubMed1985

Proteolytic self-digestion of bovine erythrocyte membranes.

"Self-digestion" of bovine erythrocyte membrane proteins was studied in isolated membrane preparations during prolonged incubation at 37 C. Protease activities associated with the membrane result in progressive degradation of all main erythrocyte membrane proteins, in particular spectrin and Band 3, and formation of lower molecular weight products which have been tentatively assigned to parent molecules. Membrane protein "self-digestion" occurs in a broad pH range (2-11), is inhibited by increased ionic strength and by inhibitors of metalloproteases, cysteine and serine proteases, and activated by low concentrations of SDS. "Self-digestion" also takes place in NaOH-stripped erythrocyte membranes. The activity of a protease involved in the "self-digestion", of apparent molecular weight of about 35,000, was renatured after SDS-polyacrylamide gel electrophoresis of erythrocyte membrane proteins.

Animals↗