PMID- 32119860
OWN - NLM
STAT- MEDLINE
DCOM- 20200820
LR  - 20200820
IS  - 1879-2642 (Electronic)
IS  - 0005-2736 (Linking)
VI  - 1862
IP  - 6
DP  - 2020 Jun 1
TI  - Non-oxidative band-3 clustering agents cause the externalization of 
      phosphatidylserine on erythrocyte surfaces by a calcium-independent mechanism.
PG  - 183231
LID - S0005-2736(20)30056-0 [pii]
LID - 10.1016/j.bbamem.2020.183231 [doi]
AB  - Aging of red blood cells (RBCs) is associated with alteration in a wide range of 
      RBC features, occurring each on its own timescale. A number of these changes are 
      interrelated and initiate a cascade of biochemical and structural 
      transformations, including band-3 clustering and phosphatidylserine (PS) 
      externalization. Using specific band-3 clustering agents (acridine orange (AO) 
      and ZnCl(2)), we examined whether treatment of RBCs with these agents may affects 
      PS externalization and whether this process is Ca(2+)-dependent. RBCs were 
      isolated from the blood of eight healthy donors upon obtaining their informed 
      consent. The suspension was supplemented with increasing concentrations of AO or 
      ZnCl(2) (from 0.5 to 2.0 mM) and incubated at 25  degrees C for 60 min. To detect PS at 
      the RBC surface, we used allophycocyanin-conjugated recombinant human Annexin V. 
      We demonstrated, that treatment of RBCs with both clustering agents caused an 
      elevation in the percent of cells positively labeled by Annexin-V (RBC(PS)), and 
      that this value was not dependent on the presence of calcium in the buffer: RBCs 
      treated with AO in the presence of either EDTA, EGTA or calcium exhibited similar 
      percentage of RBC(PS). Moreover, the active influx of Zn(2+) into RBCs induced by 
      their co-incubation with both ZnCl(2) and A23187 did not increase the percent of 
      RBC(PS) as compared to RBCs incubated with ZnCl(2) alone. Taken together, these 
      results demonstrate that the band-3 clustering agents (AO or ZnCl(2)) induce PS 
      externalization in a Ca(2+) independent manner, and we hereby suggest a possible 
      scenario for this phenomenon.
CI  - Copyright (c) 2020 Elsevier B.V. All rights reserved.
FAU - Koshkaryev, Alexander
AU  - Koshkaryev A
AD  - Biochemistry Department, The Faculty of Medicine, The Hebrew University of 
      Jerusalem, Israel.
FAU - Livshits, Leonid
AU  - Livshits L
AD  - Red Blood Cell Research Group, Institute of Veterinary Physiology, Vetsuisse 
      Faculty, University of Zurich, Zurich, Switzerland.
FAU - Pajic-Lijakovic, Ivana
AU  - Pajic-Lijakovic I
AD  - Department of Chemical Engineering, University of Belgrade, Serbia.
FAU - Gural, Alexander
AU  - Gural A
AD  - Department of Hematology, Hadassah Hebrew University Medical Center, Jerusalem, 
      Israel.
FAU - Barshtein, Gregory
AU  - Barshtein G
AD  - Biochemistry Department, The Faculty of Medicine, The Hebrew University of 
      Jerusalem, Israel. Electronic address: gregoryba@ekmd.huji.ac.il.
FAU - Yedgar, Saul
AU  - Yedgar S
AD  - Biochemistry Department, The Faculty of Medicine, The Hebrew University of 
      Jerusalem, Israel.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20200228
PL  - Netherlands
TA  - Biochim Biophys Acta Biomembr
JT  - Biochimica et biophysica acta. Biomembranes
JID - 101731713
RN  - 0 (Anion Exchange Protein 1, Erythrocyte)
RN  - 0 (Annexin A5)
RN  - 0 (Chlorides)
RN  - 0 (Phosphatidylserines)
RN  - 0 (SLC4A1 protein, human)
RN  - 0 (Zinc Compounds)
RN  - 86Q357L16B (zinc chloride)
RN  - F30N4O6XVV (Acridine Orange)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Acridine Orange/pharmacology
MH  - Anion Exchange Protein 1, Erythrocyte/*metabolism
MH  - Annexin A5/metabolism
MH  - Calcium/pharmacology
MH  - Cellular Senescence
MH  - Chlorides/pharmacology
MH  - *Cluster Analysis
MH  - Erythrocytes/*cytology
MH  - Humans
MH  - Phosphatidylserines/*metabolism
MH  - Zinc Compounds/pharmacology
OTO - NOTNLM
OT  - Acridine orange, ZnCl(2)
OT  - Band-3 aggregation
OT  - Erythrocytes
OT  - Phosphatidylserine externalization
COIS- Declaration of competing interest The authors declare that they have no known 
      competing financial interests or personal relationships that could have appeared 
      to influence the work reported in this paper.
EDAT- 2020/03/03 06:00
MHDA- 2020/08/21 06:00
CRDT- 2020/03/03 06:00
PHST- 2019/10/29 00:00 [received]
PHST- 2020/02/11 00:00 [revised]
PHST- 2020/02/14 00:00 [accepted]
PHST- 2020/03/03 06:00 [pubmed]
PHST- 2020/08/21 06:00 [medline]
PHST- 2020/03/03 06:00 [entrez]
AID - S0005-2736(20)30056-0 [pii]
AID - 10.1016/j.bbamem.2020.183231 [doi]
PST - ppublish
SO  - Biochim Biophys Acta Biomembr. 2020 Jun 1;1862(6):183231. doi: 
      10.1016/j.bbamem.2020.183231. Epub 2020 Feb 28.