O2Transfer Kinetics of Red Blood Cells Determined by Stopped-Flow Technique

Author(s):  
K. Yamaguchi ◽  
C. Hook ◽  
P. Scheid ◽  
J. Piiper
1989 ◽  
Vol 261 (3) ◽  
pp. 1009-1013 ◽  
Author(s):  
T Brittain ◽  
R Simpson

The processes of O2 uptake and release by the three embryonic haemoglobins contained within early mouse embryonic red blood cells have been studied using dual-wavelength stopped-flow kinetic spectroscopy. The rate of O2 uptake in the pseudo-spherical, nucleated, embryonic red blood cells exhibits a greater than first-order dependence on O2 concentration. The time courses for the release from the red blood cells into dithionite-containing solutions tends towards a limiting rate at high dithionite concentrations. The rates of both the uptake and release processes observed in the embryonic cells are compared with those previously seen for adult mouse red blood cells. A new mathematical model is described which accurately simulates both uptake and release experimental data for the nucleated embryonic red blood cells.


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)


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.


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

1969 ◽  
Vol 23 (3) ◽  
pp. 475-491 ◽  
Author(s):  
George H. Weiss ◽  
Gershom Zajicek

1986 ◽  
Vol 125 (1) ◽  
pp. 15-27
Author(s):  
D. Nguyen Phu ◽  
K. Yamaguchi ◽  
P. Scheid ◽  
J. Piiper

The specific conductance (G) for O2 transfer by red blood cells (RBCs) of chicken and muscovy duck was measured using the experimental (stopped-flow) and analytical techniques (RBC model) previously applied to human RBC (Yamaguchi, Nguyen Phu, Scheid & Piiper, 1985). Avian RBCs behaved similarly to human RBCs: G values were of similar magnitude; G for O2 uptake decreased with time and increasing O2 saturation; G for O2 release at high levels of dithionite decreased slightly with decreasing O2 saturation; G for O2 release was higher than G for O2 uptake. The deoxygenation kinetics of oxyhaemoglobin in solution was similar for both avian species. The G measured for O2 release at high dithionite concentration, considered to represent a good approximation to intra-erythrocyte O2 diffusion conductance, averaged (in mmol min-1 Torr-1 ml-1 RBC) 0.33 for chicken and 0.25 for duck (at 41 degrees C, pH of the suspension = 7.5, O2 saturation range 0.4-0.8). These species differences can be explained by differences in cell size, the RBC volume averaging 104 micron3 in the chicken and 155 micron3 in the duck. Compared with human RBCs, the G estimates for avian RBCs are somewhat smaller than would be predicted from size differences, which can be explained by the discoid shape of mammalian RBCs which constitutes an advantage compared with the ovoid avian RBC.


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