Anion exchange through band 3 protein in canine leishmaniasis at different stages of disease

2017 ◽  
Vol 469 (5-6) ◽  
pp. 713-724 ◽  
Author(s):  
Rossana Morabito ◽  
Alessia Remigante ◽  
Mauro Cavallaro ◽  
Alessandro Taormina ◽  
Giuseppina La Spada ◽  
...  
Keyword(s):  
Anion Exchange ◽  
Band 3 ◽  
Canine Leishmaniasis ◽  
Band 3 Protein

scholarly journals Melatonin Protects Band 3 Protein in Human Erythrocytes against H2O2-Induced Oxidative Stress

Molecules ◽  
2019 ◽  
Vol 24 (15) ◽  
pp. 2741 ◽  
Author(s):  
Rossana Morabito ◽  
Alessia Remigante ◽  
Angela Marino
Keyword(s):  
Oxidative Stress ◽  
Lipid Peroxidation ◽  
Protein Expression ◽  
Beneficial Effect ◽  
Anion Exchange ◽  
Rate Constant ◽  
Cell Shape ◽  
Band 3 ◽  
Band 3 Protein ◽  
Expression Levels

The beneficial effect of Melatonin (Mel), recognized as an anti-inflammatory and antioxidant compound, has been already proven to prevent oxidative stress-induced damage associated to lipid peroxidation. As previous studies modeled the impact of oxidative stress on Band 3 protein, an anion exchanger that is essential to erythrocytes homeostasis, by applying H2O2 at not hemolytic concentrations and not producing lipid peroxidation, the aim of the present work was to evaluate the possible antioxidant effect of pharmacological doses of Mel on Band 3 protein anion exchange capability. The experiments have been performed on human erythrocytes exposed to 300 μM H2O2-induced oxidative stress. To this end, oxidative damage has been verified by monitoring the rate constant for SO4= uptake through Band 3 protein. Expression levels of this protein Mel doses lower than 100 µM have also been excluded due to lipid peroxidation, Band 3 protein expression levels, and cell shape alterations, confirming a pro-oxidant action of Mel at certain doses. On the other hand, 100 µM Mel, not provoking lipid peroxidation, restored the rate constant for SO4= uptake, Band 3 protein expression levels, and H2O2-induced cell shape alterations. Such an effect was confirmed by abolishing the endogenous erythrocytes antioxidant system. Therefore, the present findings show the antioxidant power of Mel at pharmacological concentrations in an in vitro model of oxidative stress not associated to lipid peroxidation, thereby confirming Band 3 protein anion exchange capability measurement as a suitable model to prove the beneficial effect of Mel and support the use of this compound in oxidative stress-related diseases affecting Band 3 protein.

Impact of acute inflammation on Band 3 protein anion exchange capability in human erythrocytes

2020 ◽  
pp. 1-7 ◽  
Author(s):  
Rossana Morabito ◽  
Alessia Remigante ◽  
Marika Cordaro ◽  
Vincenzo Trichilo ◽  
Saverio Loddo ◽  
...  
Keyword(s):  
Anion Exchange ◽  
Acute Inflammation ◽  
Band 3 ◽  
Human Erythrocytes ◽  
Band 3 Protein

scholarly journals Catecholamine-induced transport systems in trout erythrocyte. Na+/H+ countertransport or NaCl cotransport?

1986 ◽  
Vol 87 (4) ◽  
pp. 551-566 ◽  
Author(s):  
F Borgese ◽  
F Garcia-Romeu ◽  
R Motais
Keyword(s):  
Anion Exchange ◽  
Band 3 ◽  
Cell Swelling ◽  
Transport Systems ◽  
Band 3 Protein ◽  
Free Medium ◽  
Exchange System ◽  
Nucleated Red Blood Cells ◽  
Nacl Cotransport ◽  
Na Uptake

It has previously been shown (Baroin, A., F. Garcia-Romeu, T. Lamarre, and R. Motais. 1984a, b. Journal of Physiology. 350:137, 356:21; Mahé, Y., F. Garcia-Romeu, and R. Motais. 1985. European Journal of Pharmacology. 116:199) that the addition of catecholamines to an isotonic suspension of nucleated red blood cells of the rainbow trout first stimulates a cAMP-dependent, amiloride-sensitive Na+/H+ exchange. This stimulation seems to be transient. It is followed by a more permanent activation of a coupled entry of Na+ and Cl-, which is inhibited by amiloride but also by inhibitors of band 3 protein (DIDS, furosemide, niflumic acid). The coupled entry of Na+ and Cl- could therefore result from the parallel and simultaneous exchange of Na+out for H+in (via the cAMP-dependent Na+/H+ antiporter) and Cl- out for HCO3- in (via the anion exchange system located in band 3 protein). However, in view of the following arguments, it had been proposed that NaCl uptake does not proceed by the double-exchanger system but via an NaCl cotransport: (a) Na+ entry requires Cl- as anion (in NO3- medium, the Na uptake is strongly inhibited, whereas NO3- is an extremely effective substitute for Cl- in the anion exchange system); (b) Na uptake is not significantly affected by the presence of HCO3- in the suspension medium despite the fact that in red cells, Cl-/HCO3- exchange occurs more readily than the exchanges of Cl- for basic equivalents in a theoretically CO2-free medium (the so-called Cl-/OH- exchanges). The purpose of the present paper was a reassessment of the two models by using monensin, an ionophore allowing Na+/H+ exchange. From this study, it appears that NaCl entry results from the simultaneous functioning of the Na+/H+ antiporter and the anion exchange system. The apparent Cl dependence is explained by the fact that, in these erythrocytes, NO3- clearly inhibits the turnover rate of the Na+/H+ antiporter. As Na+/H+ exchange is the driving component in the salt uptake process, this inhibition explains the Cl requirement for Na entry. The lack of stimulation of cell swelling by bicarbonate is explained by the fact that the rate of anion exchange in a CO2-free medium (Cl-/OH- exchange) is roughly equivalent to that of Na+/H+ exchange and thus in practice is not limiting to the net influx of NaCl through the two exchangers.(ABSTRACT TRUNCATED AT 400 WORDS)

scholarly journals Natural Antioxidants Beneficial Effects on Anion Exchange through Band 3 Protein in Human Erythrocytes

Antioxidants ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 25 ◽  
Author(s):  
Alessia Remigante ◽  
Rossana Morabito ◽  
Angela Marino
Keyword(s):  
Anion Exchange ◽  
Natural Antioxidants ◽  
Band 3 ◽  
Erythrocyte Membranes ◽  
Band 3 Protein ◽  
Ion Distribution ◽  
Beneficial Effects ◽  
Cross Links

Band 3 protein (B3p) exchanging Cl− and HCO3− through erythrocyte membranes is responsible for acid balance, ion distribution and gas exchange, thus accounting for homeostasis of both erythrocytes and entire organisms. Moreover, since B3p cross links with the cytoskeleton and the proteins underlying the erythrocyte membrane, its function also impacts cell shape and deformability, essential to adaptation of erythrocyte size to capillaries for pulmonary circulation. As growing attention has been directed toward this protein in recent years, the present review was conceived to report the most recent knowledge regarding B3p, with specific regard to its anion exchange capability under in vitro oxidative conditions. Most importantly, the role of natural antioxidants, i.e., curcumin, melatonin and Mg2+, in preventing detrimental oxidant effects on B3p is considered.

Pre-steady state transport by erythrocyte band 3 protein: uphill countertransport induced by the impermeant inhibitor H2DIDS

1998 ◽  
Vol 76 (5) ◽  
pp. 807-813 ◽  
Author(s):  
Michael L Jennings ◽  
Jonathan Whitlock ◽  
Anjali Shinde
Keyword(s):  
Steady State ◽  
Anion Exchange ◽  
Sodium Phosphate ◽  
Band 3 ◽  
Catalytic Cycle ◽  
Band 3 Protein ◽  
Molar Excess ◽  
Transport Inhibitor ◽  
Ping Pong

Pre-steady state Cl- efflux experiments have been performed to test directly the idea that the transport inhibitor H2DIDS (4,4'-diisothiocyanatodihydrostilbene-2,2'-disulfonate) binds preferentially to the outward-facing state of the transporter. Cells were equilibrated with a medium consisting of 150 mM sodium phosphate, pH 6.2, N2 atmosphere, and 80-250 µM 36Cl-. Addition of H2DIDS (10-fold molar excess compared with band 3) induces a transient efflux of Cl-, as expected if H2DIDS binds more tightly to outward-facing than to inward-facing states. The size of the H2DIDS-induced efflux depends on the Cl- concentration and is about 700 000 ions per cell at the highest concentrations tested. The size of the transient efflux is larger than would be expected if the catalytic cycle for anion exchange involved one pair of exchanging anions per band 3 dimer. These results are completely consistent with a ping-pong mechanism of anion exchange in which the catalytic cycle consists of one pair of exchanging anions per subunit of the band 3 dimer.Key words: anion exchange, erythrocyte, stilbenedisulfonate, chloride.

Hydroxyurea Induced Expression of Erythrocyte Anion Exchange Protein [AE1] and Gamma Globin in Cultured Erythroid Progenitor Cells.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3536-3536
Author(s):  
Gloria A. Green ◽  
Beau R. Braden ◽  
Obianuju Mba ◽  
Stacy A. Chivira ◽  
Laleh Ramezani ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Anion Exchange ◽  
Sickle Cell ◽  
Band 3 ◽  
Band 3 Protein ◽  
Erythroid Progenitors ◽  
Gamma Globin ◽  
Anion Exchange Protein ◽  
Reactive Mechanism ◽  
Cells Cultured

Abstract Background: Hydroxyurea (HU) an S-phase specific cytotoxic agent has been used for the treatment of patients with sickle cell anemia and beta-thalassemia. The clinical efficacy of HU is due primarily to increases in fetal hemoglobin (HbF) levels. HU increases the %HbF and the %F cells. The HU reactive mechanism(s) in erythroid cells, however, have not been clearly defined. Patients receiving HU therapy develop subpopulations of macrocytic erythrocytes. Our previous studies demonstrate that sickle cell patients treated with low dose HU develop subpopulations of RBCs that express greater relative levels of the erythrocyte anion exchange protein (AE1) per cell as compared with untreated individuals. The frequency of cells expressing greater levels of (AE1) per cell was increased in each serial blood sample. We propose that part of the HU reactive mechanism will include the upmodulation of non-gamma globin erythroid proteins that contribute to the macrocytic structures. As part of our investigation of the development of RBCs expressing increased band 3 protein per cell, we examined the possibility that HU induced AE1 synthesis can be detected in vitro using cultured erythroid progenitors. Methods: HU induced protein synthesis was investigated as a function of HU concentration. Erythroid progenitors were cultured in serum free semisolid media containing different concentrations of HU [0–40 micromolar]. BFU-E were scored and harvested after 15 days in culture, then assayed. BFU-E derived cells were first labeled with monoclonal anti-band 3 antibody. The change in the frequency of cells positive for band 3 protein was determined by flow cytometry. BFU-E derived cells were subsequently labeled with anti-spectrin antibody plus monoclonal anti-band 3 antibody, then assayed. For the next set of experiments, BFU-E cultured in increasing concentrations of HU were assayed for the presence of band 3 protein and gamma globin. BFU-E cells were fixed permeabilized, and then labeled with tricolor-conjugated-anti-gamma globin. These cells were labeled with monoclonal anti-band 3 and PE-labeled anti-mouse antibody, assayed by flow cytometry. Results: Results show that the frequency of cells positive for band 3 protein [AE1] was increased at increasing concentrations of HU as compared to controls. The band-3 upmodulation appears to plateau at 12.5–20 micromolar HU. Band 3 protein derived from BFUE was confirmed by Western blot analysis. The frequency of cells positive for both band 3 and spectrin was similarly increased in cells cultured in the presence of increasing concentrations of HU. In separate studies, cells cultured in HU were assayed for the expression of both gamma globin and AE1 by flow cytometry. Results show a 2–3-fold increase in the % band 3 [AE1] plus gamma globin positive cells in HU [5–40 micromolar] treated cells as compared to untreated controls. Conclusions: These results demonstrate that HU induces the expression of band 3 and gamma globin in cultured erythroid progenitors. These findings suggest that part of the mechanism of HU action in erythroid cells involves the induction of erythroid structural proteins concordant with the induction of gamma globin.

scholarly journals d-Galactose Decreases Anion Exchange Capability through Band 3 Protein in Human Erythrocytes

Antioxidants ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 689
Author(s):  
Alessia Remigante ◽  
Rossana Morabito ◽  
Sara Spinelli ◽  
Vincenzo Trichilo ◽  
Saverio Loddo ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Lipid Peroxidation ◽  
Anion Exchange ◽  
Band 3 ◽  
Human Erythrocytes ◽  
Band 3 Protein ◽  
Expression Levels ◽  
Vitro Effect ◽  
The Impact

d-Galactose (d-Gal), when abnormally accumulated in the plasma, results in oxidative stress production, and may alter the homeostasis of erythrocytes, which are particularly exposed to oxidants driven by the blood stream. In the present investigation, the effect of d-Gal (0.1 and 10 mM, for 3 and 24 h incubation), known to induce oxidative stress, has been assayed on human erythrocytes by determining the rate constant of SO42− uptake through the anion exchanger Band 3 protein (B3p), essential to erythrocytes homeostasis. Moreover, lipid peroxidation, membrane sulfhydryl groups oxidation, glycated hemoglobin (% A1c), methemoglobin levels (% MetHb), and expression levels of B3p have been verified. Our results show that d-Gal reduces anion exchange capability of B3p, involving neither lipid peroxidation, nor oxidation of sulfhydryl membrane groups, nor MetHb formation, nor altered expression levels of B3p. d-Gal-induced %A1c, known to crosslink with B3p, could be responsible for rate of anion exchange alteration. The present findings confirm that erythrocytes are a suitable model to study the impact of high sugar concentrations on cell homeostasis; show the first in vitro effect of d-Gal on B3p, contributing to the understanding of mechanisms underlying an in vitro model of aging; demonstrate that the first impact of d-Gal on B3p is mediated by early Hb glycation, rather than by oxidative stress, which may be involved on a later stage, possibly adding more knowledge about the consequences of d-Gal accumulation.

scholarly journals Effect of cadmium on anion exchange capability through Band 3 protein in human erythrocytes

2018 ◽  
Vol 91 (1) ◽  
Author(s):  
Rossana Morabito ◽  
Alessia Remigante ◽  
Benedetta Arcuri ◽  
Angela Marino ◽  
Marco Giammanco ◽  
...  
Keyword(s):  
Protein Expression ◽  
Anion Exchange ◽  
Rate Constant ◽  
Band 3 ◽  
Human Erythrocytes ◽  
Band 3 Protein ◽  
Toxic Heavy Metal ◽  
Expression Levels ◽  
Sh Groups ◽  
The Impact

The efficiency of Band 3 protein, mediating HCO3-/Cl- exchange across erythrocytes membrane, is reduced by oxidative stress. The aim of the present study was to verify whether Band 3 protein efficiency is compromised by treatment with Cadmium (Cd2+), an extremely toxic heavy metal known to interfere with antioxidant enzymes, energy metabolism, gene expression and cell membranes. To this end, the rate constant for SO4= uptake through Band 3 protein (accounting for velocity of anion exchange) was measured along with membrane –SH groups, Malonyldialdehyde (MDA) and Band 3 protein expression levels in Cd2+ -treated human erythrocytes (300 µM, 1 mM). Our results show that Cd2+ reduced the rate constant for SO4= uptake, with a significant increase in MDA levels at both concentrations and with a reduction in –SH groups observed after 1 mM Cd2+ treatment, whereas Band 3 protein expression levels were unchanged in both conditions. In conclusion: i) Cd2+ reduces Band 3 protein efficiency via different mechanisms depending on metal concentration and with unchanged expression levels; ii) the assessment of Band 3 protein anion exchange capability is a good tool to assay the impact of heavy metals on cell homeostasis and, possibly, useful for diagnosis and monitoring of devalopment of Cd2+ toxicity-related pathologies.

scholarly journals High Glucose Concentrations Affect Band 3 Protein in Human Erythrocytes

Antioxidants ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 365 ◽  
Author(s):  
Rossana Morabito ◽  
Alessia Remigante ◽  
Sara Spinelli ◽  
Giulia Vitale ◽  
Vincenzo Trichilo ◽  
...  
Keyword(s):  
Anion Exchange ◽  
Rate Constant ◽  
High Glucose ◽  
Thiobarbituric Acid ◽  
Band 3 ◽  
Sulfhydryl Groups ◽  
Band 3 Protein ◽  
Expression Levels ◽  
The Impact

Hyperglycemia is considered a threat for cell homeostasis, as it is associated to oxidative stress (OS). As erythrocytes are continuously exposed to OS, this study was conceived to verify the impact of either diabetic conditions attested to by glycated hemoglobin (Hb) levels (>6.5% or higher) or treatment with high glucose (15–35 mM, for 24 h) on erythrocyte homeostasis. To this aim, anion exchange capability through the Band 3 protein (B3p) was monitored by the rate constant for SO42− uptake. Thiobarbituric acid reactive species (TBARS), membrane sulfhydryl groups mostly belonging to B3p, glutathione reduced (GSH) levels, and B3p expression levels were also evaluated. The rate constant for SO42− uptake (0.063 ± 0.001 min−1, 16 min in healthy volunteers) was accelerated in erythrocytes from diabetic volunteers (0.113 ± 0.001 min−1, 9 min) and after exposure to high glucose (0.129 ± 0.001in−1, 7 min), but only in diabetic volunteers was there an increase in TBARS levels and oxidation of membrane sulfhydryl groups, and a decrease in both GSH and B3p expression levels was observed. A combined effect due to the glycated Hb and OS may explain what was observed in diabetic erythrocytes, while in in vitro hyperglycemia, early OS could explain B3p anion exchange capability alterations as proven by the use of melatonin. Finally, measurement of B3p anion exchange capability is a suitable tool to monitor the impact of hyperglycemia on erythrocytes homeostasis, being the first line of high glucose impact before Hb glycation. Melatonin may be useful to counteract hyperglycemia-induced OS at the B3p level.

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