Non-oxidative band-3 clustering agents cause the externalization of phosphatidylserine on erythrocyte surfaces by a calcium-independent mechanism

Koshkaryev, Alexander; Livshits, Leonid; Pajić‐Lijaković, Ivana; Gural, Alexander; Barshtein, Gregory; Yedgar, Saul

doi:10.1016/j.bbamem.2020.183231

Cited by 10 publications

(13 citation statements)

References 84 publications

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Section: Discussionmentioning

confidence: 68%

“…These previous reports using PMA and LPA 2 , 35 failed to show a clear correlation between red cells with elevations in Ca 2+ and PS exposure. They also suggested that PS exposure could not occur in the absence of extracellular Ca 2+ 2 , unlike the present findings. There is an important caveat here in that the high Ca 2+ affinity of the fluorophores used (fluo3/4) is such that cells would show positive for Ca 2+ at low submicrolar concentrations which may be insufficient to cause Ca 2+ -induced PS scrambling, which occurs at an EC 50 of about 1 μM 6 , 18 , 37 .…”

Section: Discussionmentioning

confidence: 68%

“…Rather there must be some alternative mechanism of action of Yoda1. Previously, Yoda1 has been shown to activate protein kinases in other tissues 34 , and there is also strong evidence of a role for protein kinase C (PKC) in PS exposure 2 , 35 , 36 , either via Ca 2+ entry through cation channels or via a Ca 2+ -independent action. The effect of Yoda1 in sickle cells was therefore examined in combination with inhibitors of PKC, chelerytherine chloride which inhibits the active phosphorylation site of PKC, and calphostin C, which is an irreversible inhibitor of the diacylglyceraldehye-binding site.…”

Section: Resultsmentioning

confidence: 99%

“…7b), indicating an additional effect of intracellular Ca 2+ independent of PKC, although it is also possible that PKC inhibition used here was incompleteat the concentration of chelerytherine chloride. Previous work using phorbol myristate acetate (PMA), lysophosphatidic acid (LPA) and the Ca 2+ ionophore A23187 together with chelerytherine chloride and calphostin C has also produced evidence for PKC-mediated PS exposure in normal and sickle cells, through both Ca 2+ -dependent and Ca 2+ -independent mechanisms 2,35,36,39 . The Ca 2+ -dependent effect of PKC could be mediated via Ca 2+ entry, with the participation of w-agatoxin-TKsensitive, Cav2.1-like, Ca 2+ channels or possibly the non-selection cation channel 20,40 .…”

Section: Discussionmentioning

confidence: 99%

“…Normally less than 1% red cells are positive for externalised PS. Notwithstanding, increased PS exposure does occur in healthy red cells during their normal ageing process 2 – 4 . In addition, however, red cells from patients with sickle cell anaemia (SCA) show less PS asymmetry than those from normal individuals, such that a high, but variable, proportion (c.1–10%), are positive for exposed PS 5 , 6 .…”

Section: Introductionmentioning

confidence: 99%

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Yoda1 and phosphatidylserine exposure in red cells from patients with sickle cell anaemia

Wadud

Hannemann

Rees

et al. 2020

Sci Rep

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Phosphatidylserine (PS) exposure is increased in red cells from sickle cell anaemia (SCA) patients. Externalised PS is prothrombotic and attractive to phagocytes and activated endothelial cells and thus contributes to the anaemic and ischaemic complications of SCA. The mechanism of PS exposure remains uncertain but it can follow increased intracellular Ca2+ concentration ([Ca2+]i). Normally, [Ca2+]i is maintained at very low levels but in sickle cells, Ca2+ permeability is increased, especially following deoxygenation and sickling, mediated by a pathway sometimes called Psickle. The molecular identity of Psickle is also unclear but recent work has implicated the mechanosensitive channel, PIEZO1. We used Yoda1, an PIEZO1 agonist, to investigate its role in sickle cells. Yoda1 caused an increase in [Ca2+]i and PS exposure, which was inhibited by its antagonist Dooku1 and the PIEZO1 inhibitor GsMTx4, consistent with functional PIEZO1. However, PS exposure did not necessitate an increase in [Ca2+]i. Two PKC inhibitors were also tested, chelerytherine chloride and calphostin C. Both reduced PS exposure whilst chelerytherine chloride also reduced Yoda1-induced increases in [Ca2+]i. Findings are therefore consistent with the presence of PIEZO1 in sickle cells, able to mediate Ca2+ entry but that PKC was also involved in both Ca2+ entry and PS exposure.

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Section: Discussionmentioning

confidence: 68%

Section: Discussionmentioning

confidence: 68%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Yoda1 and phosphatidylserine exposure in red cells from patients with sickle cell anaemia

Wadud

Hannemann

Rees

et al. 2020

Sci Rep

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d‐Galactose induced early aging in human erythrocytes: Role of band 3 protein

Spinelli

Trichilo²,

Loddo³

et al. 2021

Journal Cellular Physiology

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Aging, a time-dependent multifaceted process, affects both cell structure and function and involves oxidative stress as well as glycation. The present investigation focuses on the role of the band 3 protein (B3p), an anion exchanger essential to red cells homeostasis, in a D-galactose (D-Gal)-induced aging model. Anion exchange capability, measured by the rate constant of SO₄² − uptake through B3p, levels of lipid peroxidation, oxidation of membrane sulfhydryl groups, B3p expression, methemoglobin, glycated hemoglobin (Hb), and the reduced glutathione/oxidized glutathione ratio were determined after exposure of human erythrocytes to 25, 35, 50, and 100 mmol/L D-Gal for 24 h. Our results show that: (i) in vitro application of D-Gal is useful to model early aging in human erythrocytes; (ii) assessment of B3p ion transport function is a sensitive tool to monitor aging development; (iii) D-Gal leads to Hb glycation and produces substantial changes on the endogenous antioxidant system; (iv) the impact of aging on B3p function proceeds through steps, first involving Hb glycation and then oxidative events at the membrane level. These findings offer a useful tool to understand the mechanisms of aging in human erythrocytes and propose B3p as a possible target for new therapeutic strategies to counteract age-related disturbances.

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