Oxidative Damage Does Not Alter Membrane Phospholipid Asymmetry in Human Erythrocytes†

Biochemistry ◽  
1997 ◽  
Vol 36 (22) ◽  
pp. 6768-6776 ◽  
Author(s):  
Kitty de Jong ◽  
Danielle Geldwerth ◽  
Frans A. Kuypers
Keyword(s):  
Oxidative Damage ◽  
Human Erythrocytes ◽  
Membrane Phospholipid ◽  
Phospholipid Asymmetry

scholarly journals Monitoring of membrane phospholipid scrambling in human erythrocytes and K562 cells with FM1-43 — a comparison with annexin V-FITC

2014 ◽  
Vol 19 (2) ◽  
Author(s):  
Anna Wróbel ◽  
Małgorzata Bobrowska-Hägerstrand ◽  
Christer Lindqvist ◽  
Henry Hägerstrand
Keyword(s):  
Annexin V ◽  
K562 Cells ◽  
Human Erythrocytes ◽  
Membrane Phospholipid ◽  
Ionophore A23187 ◽  
Asymmetric Membrane ◽  
Phospholipid Asymmetry ◽  
Styryl Dye ◽  
Phospholipid Scrambling ◽  
Phospholipid Distribution

AbstractThe styryl dye FM1-43 becomes highly fluorescent upon binding to cell membranes. The breakdown of membrane phospholipid asymmetry in ionophore-stimulated T-lymphocytes further increases this fluorescence [Zweifach, 2000]. In this study, the capacity of FM1-43 to monitor membrane phospholipid scrambling was explored using flow cytometry in human erythrocytes and human erythrocyte progenitor K562 cells. The Ca2+-dependent phosphatidylserine-specific probe annexin V-FITC was used for comparison. The presented data show that the loss of phospholipid asymmetry that could be induced in human erythrocytes by elevated intracellular Ca2+ or by structurally different membrane intercalated amphiphilic compounds increases the FM1-43 fluorescence two- to fivefold. The profile of FM1-43 fluorescence for various treatments resembles that of phosphatidylserine exposure reported by annexin V-FITC. FM1-43 detected the onset of scrambling more efficiently than annexin V-FITC. The amphiphile-induced scrambling was shown to be a Ca2+-independent process. Monitoring of scrambling in K562 cells caused by NEM-induced Ca2+-release from intracellular stores and by Ca2+ and ionophore A23187 treatment showed that the increase in FM1-43 fluorescence correlated well with the number of annexin V-FITC-detected phosphatidylserine-positive cells. The results presented here show the usefulness of FM1-43 as a Ca2+-independent marker of dissipation in asymmetric membrane phospholipid distribution induced by various stimuli in both nucleated and non-nucleated cells.

scholarly journals The complex of phosphatidylinositol 4,5-bisphosphate and calcium ions is not responsible for Ca(2+)-induced loss of phospholipid asymmetry in the human erythrocyte: a study in Scott syndrome, a disorder of calcium- induced phospholipid scrambling

Blood ◽  
1995 ◽  
Vol 86 (5) ◽  
pp. 1983-1991 ◽  
Author(s):  
EM Bevers ◽  
T Wiedmer ◽  
P Comfurius ◽  
J Zhao ◽  
EF Smeets ◽  
...  
Keyword(s):  
Lipid Vesicles ◽  
Human Erythrocytes ◽  
Mechanism Analysis ◽  
Membrane Phospholipids ◽  
Mole Percent ◽  
Cellular Processes ◽  
Phospholipid Asymmetry ◽  
Dependent Cell ◽  
Normal Human ◽  
State Concentration

Elevation of cytoplasmic Ca2+ levels in human erythrocytes induces a progressive loss of membrane phospholipid asymmetry, a process that is impaired in erythrocytes from a patient with Scott syndrome. We show here that porcine erythrocytes are similarly incapable of Ca(2+)- induced redistribution of membrane phospholipids. Because a complex of phosphatidylinositol 4,5-bisphosphate (PIP2) and Ca2+ has been proposed as the mediator of enhanced transbilayer movement of lipids (J Biol Chem 269:6347,1994), these cell systems offer a unique opportunity for testing this mechanism. Analysis of both total PIP2 content and the metabolic-resistant pool of PIP2 that remains after incubation with Ca2+ ionophore showed no appreciable differences between normal and Scott erythrocytes. Moreover, porcine erythrocytes were found to have slightly higher levels of both total and metabolic-resistant PIP2 in comparison with normal human erythrocytes. Although loading of normal erythrocytes with exogenously added PIP2 gave rise to a Ca(2+)-induced increase in prothrombinase activity and apparent transbilayer movement of nitrobenzoxadiazolyl (NBD)-phospholipids, these PIP2-loaded cells were also found to undergo progressive Ca(2+)-dependent cell lysis, which seriously hampers interpretation of these data. Moreover, loading Scott cells with PIP2 did not abolish their impaired lipid scrambling, even in the presence of a Ca(2+)-ionophore. Finally, artificial lipid vesicles containing no PIP2 or 1 mole percent of PIP2 were indistinguishable with respect to transbilayer movement of NBD- phosphatidylcholine in the presence of Ca2+. Our findings suggest that Ca(2+)-induced redistribution of membrane phospholipids cannot simply be attributed to the steady-state concentration of PIP2, and imply that such lipid movement is regulated by other cellular processes.

Crocin protects human erythrocytes from nitrite-induced methemoglobin formation and oxidative damage

2016 ◽  
Vol 40 (12) ◽  
pp. 1320-1331 ◽  
Author(s):  
Fariheen Aisha Ansari ◽  
Shaikh Nisar Ali ◽  
Riaz Mahmood
Keyword(s):  
Oxidative Damage ◽  
Human Erythrocytes ◽  
Methemoglobin Formation
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