Comparison between Ca2+-induced scrambling of various fluorescently labelled lipid analogues in red blood cells

2002 ◽  
Vol 362 (3) ◽  
pp. 741-747 ◽  
Author(s):  
David W. C. DEKKERS ◽  
Paul COMFURIUS ◽  
Edouard M. BEVERS ◽  
Robert F. A. ZWAAL

Treatment of red blood cells with calcium and ionomycin causes activation of the lipid scramblase, a putative membrane protein catalysing flip-flop of (phospho)lipids. Various fluorescent 1-oleoyl-2-[6(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino] caproyl (C6-NBD) analogues were tested for transbilayer movement across the plasma membrane of red blood cells. Among these phospholipid analogues were phosphatidylgalactose, phosphatidylmaltose and phosphatidylmaltotriose, which were obtained from C6-NBD-phosphatidylcholine by phospholipase D-catalysed transphosphatidylation. The inward movement after the onset of scrambling was monitored by extraction of the non-internalized probe with BSA. We demonstrate that both the amino group and the size of the headgroup determine the kinetics of lipid scrambling, and that lipids with a ceramide backbone migrate much more slowly than glycerophospholipids with the same headgroup.

Author(s):  
Néstor J. Oviedo ◽  
Gustavo Benaim ◽  
Vincenza Cervino ◽  
Teresa Proverbio ◽  
Fulgencio Proverbio ◽  
...  

1996 ◽  
Vol 12 (4) ◽  
pp. 94-99 ◽  
Author(s):  
T. V. Stasyk ◽  
M. D. Lutsik-Kordovsky

2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Krzysztof Gwozdzinski ◽  
Anna Pieniazek ◽  
Joanna Bernasinska-Slomczewska ◽  
Joanna Brzeszczynska ◽  
Robert Irzmanski ◽  
...  

Purpose. Comprehensive cardiac rehabilitation (CCR) is a complex program aimed at improving the health status of patients with coronary artery disease (CAD), especially those who have been subjected to cardiac interventions (PCI and CABG).The aim of this study was to measure the changes in the properties of red blood cells (RBCs) in men with CAD after cardiac intervention and after participation in CCR program. Methods. In this study, we have investigated the influence of the physical training-based CCR program in 12 men with CAD, after PCI or CABG. The characteristics of RBCs including the basic morphology of RBCs, the conformational state of RBC membrane protein and hemoglobin, acetylcholinesterase activity, membrane fluidity, the osmotic fragility, and thiol concentration in membrane and in hemolysate were measured. Ascorbate concentration and reduced glutathione were also determined. The analysis was performed in men, before and after participation in CCR. The properties of RBCs were observed in connection with the exercise test, and parameters were evaluated before, immediately after, and 1 hour after the exercise test. Results. After CCR, a decrease in the mobility of erythrocyte membrane proteins was observed, which was accompanied by a decrease in lipid fluidity. In addition, immediately after the exercise test and 1 hour later, we measured a decrease in thiol level in hemolysate, but not in the plasma membrane. Unexpectedly, an increase in reduced glutathione concentration one hour after the exercise test after completing comprehensive cardiac rehabilitation was observed. Conclusion. CCR in men with CAD after cardiac intervention is connected with decreased membrane fluidity and decreased membrane protein mobility, which indicates that reduction of oxidative changes in these components occurs.


1989 ◽  
Vol 257 (1) ◽  
pp. C58-C64 ◽  
Author(s):  
K. H. Ryu ◽  
N. C. Adragna ◽  
P. K. Lauf

The kinetic parameters and transport mechanism of Na-Li exchange were studied in both low K (LK) and high K (HK) sheep red blood cells with cellular Na [( Na]i) and Li concentrations [( Li]i) adjusted by the nystatin technique (Nature New Biol. 244: 47-49, 1973 and J. Physiol. Lond. 283: 177-196, 1978). Maximum velocities (Vm) for Li fluxes and half-activation constants (K1/2) for Li and Na of the Na-Li exchanger were determined. The K1/2 values for both Li and Na appeared to be similar in both cell types, although they were about two to three times lower on the inside than on the outside of the membrane. Furthermore, the K1/2 values for Li were at least an order of magnitude smaller than those for Na, suggesting substantial affinity differences for these two cations. The Vm values for Li fluxes, on the other hand, appear to be lower in HK than in LK cells. When Na and Li fluxes were measured simultaneously, a trans stimulatory effect by Na on Li fluxes was observed. From measurements of Li influx at different concentrations of external Li and different [Na]i, the ratio of the apparent Vm to the apparent external Li affinity was calculated to be independent of [Na]i for both types of sheep red blood cells. Similar trans effects of external Na were observed on Li efflux at varying [Li]i. These results are expected for a system operating by a “ping-pong” mechanism.


1993 ◽  
Vol 265 (1) ◽  
pp. C99-C105 ◽  
Author(s):  
Z. C. Xu ◽  
P. B. Dunham ◽  
B. Dyer ◽  
R. Blostein

Na(+)-K+ pumps of red blood cells from sheep of the low-K+ (LK) phenotype undergo differentiation during circulation, manifested in part by a striking increase in sensitivity to inhibition by intracellular K+ (Ki). Pumps of red blood cells from sheep from the allelic phenotype, high K+ (HK), do not undergo this type of maturation. The hypothesis was tested that the Lp antigen, found on LK but not HK cells, is responsible for the maturation of LK pumps. Lp antigens have been shown to inhibit LK pumps because anti-Lp antibody stimulates the pumps by relieving inhibition by the antigen. Lp antigens were recently shown to be molecular entities separate from Na(+)-K+ pumps [Xu, Z.-C., P. Dunham, J. Munzer, J. Silvius, and R. Blostein. Am. J. Physiol. 263 (Cell Physiol. 32): C1007-C1014, 1992]. The test of the hypothesis was to modify the Lp antigens of immature LK red blood cells with two kinds of treatments, anti-Lp antibody and trypsinization (which cleaves Lp), and to observe the effects of these treatments on maturation of pumps during culture of the cells in vitro. Both of these treatments prevented the maturation of the kinetics of the pumps to the Ki-sensitive pattern, supporting the hypothesis that interaction of the pumps with Lp antigens is responsible for the maturation of the pumps. Strong supportive evidence came from experiments on Na(+)-K+ pumps from rat kidney delivered into immature LK sheep red blood cells by microsome fusion.(ABSTRACT TRUNCATED AT 250 WORDS)


1997 ◽  
Vol 25 (3) ◽  
pp. 492S-492S ◽  
Author(s):  
Mei M. Ho ◽  
Anna Nicolaou ◽  
Annette C. Argent ◽  
Gordon W. Stewart

2018 ◽  
Vol 3 (3) ◽  
pp. 293-304 ◽  
Author(s):  
Andra C. Dumitru ◽  
Mégane A. Poncin ◽  
Louise Conrard ◽  
Yves F. Dufrêne ◽  
Donatienne Tyteca ◽  
...  

Red blood cells present a complex cell plasma membrane architecture with submicrometric organization leading to nanomechanical heterogeneities.


1991 ◽  
Vol 97 (2) ◽  
pp. 173-193 ◽  
Author(s):  
E Delpire ◽  
P K Lauf

A detailed kinetic study of K:Cl cotransport in hyposmotically swollen low K sheep red blood cells was carried out to characterize the nature of the outwardly poised carrier. The kinetic parameters were determined from the rate of K efflux and influx under zero-K-trans conditions in red cells with cellular K altered by the nystatin method and with different extracellular K or Rb concentrations. Although apparent affinities for efflux and influx were quite similar, the maximal velocity for K efflux was approximately two times greater than for influx. Furthermore, at thermodynamic equilibrium (i.e., when the ion product of K and Cl within the cell was equal to that outside) a temperature-dependent net K efflux was observed, approaching zero only when the external product reached approximately two times the internal product. The binding order of the ions to the transporter was asymmetric, being ordered outside (Cl binding first, followed by K) and random inside. K efflux but not influx was trans-inhibited by KCl. Trans inhibition of K efflux was used to verify the order of binding outside: trans inhibition by external Cl occurred in the absence of external K, but not vice versa. Thus K:Cl cotransport is kinetically asymmetric in hyposmotically swollen low K sheep red cells.


1989 ◽  
Vol 75 (3) ◽  
pp. 371-384 ◽  
Author(s):  
K. Yamaguchi ◽  
M.L. Glass ◽  
P. Scheid ◽  
J. Piiper

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