scholarly journals Comparison of human red cell lysis by hypochlorous and hypobromous acids: insights into the mechanism of lysis

1998 ◽  
Vol 330 (1) ◽  
pp. 131-138 ◽  
Author(s):  
C. M. Margret VISSERS ◽  
C. Anitra CARR ◽  
L. P. Anna CHAPMAN
Keyword(s):  
Membrane Protein ◽  
Protein Modification ◽  
Hypochlorous Acid ◽  
Membrane Lipids ◽  
Band 3 ◽  
Cytoskeletal Proteins ◽  
Red Cells ◽  
Fluorescent Label ◽  
Human Red Blood Cells ◽  
High Concentrations

Human red blood cells are lysed by the neutrophil-derived oxidant hypochlorous acid (HOCl), although the mechanism of lysis is unknown. Hypobromous acid (HOBr), a similarly reactive oxidant, lysed red cells approx. 10-fold faster than HOCl. Therefore we compared the effects of these oxidants on thiols, membrane lipids and proteins to determine which reactions are associated with lysis. There was no difference in the loss of reduced glutathione or membrane thiols with either oxidant, but HOBr reacted more readily with membrane lipids and proteins. Bromohydrin derivatives of phospholipids and cholesterol were seen at approx. one-tenth the level of oxidant than chlorohydrins were. However, these products were detected only with high concentrations of HOCl or HOBr, which caused instant haemolysis. Membrane protein modification occurred at much lower doses of oxidant and was more closely correlated with lysis. SDS/PAGE analysis showed that band 3, the anion transport protein, was lost at the lowest dose of HOBr and at the higher concentrations of HOCl. Labelling the red cells with eosin 5-maleimide, a fluorescent label for band 3, suggested possible clustering of this protein in oxidant-exposed cells. There was also irreversible cross-linking of all the major membrane proteins; this reaction occurred more readily with HOBr. The results indicate that membrane protein modification is the reaction responsible for HOCl-mediated lysis. These effects, and particularly cross-link formation, might result in clustering of band 3 and other membrane and cytoskeletal proteins to form haemolytic pores.

scholarly journals The Uptake of Divalent Manganese Ion by Mature Normal Human Red Blood Cells

1960 ◽  
Vol 44 (2) ◽  
pp. 301-314 ◽  
Author(s):  
Robert I. Weed ◽  
Aser Rothstein
Keyword(s):  
Red Cells ◽  
Metabolic Inhibitors ◽  
Entry And Exit ◽  
Permeability Constant ◽  
Manganese Ion ◽  
Human Red Blood Cells ◽  
Metabolic Substrates ◽  
Normal Human ◽  
High Concentrations ◽  
Mn Concentration

At physiological pH and concentrations of Mn++ in excess of 5 x 10-4 M, study of the Mn++ ion movement into human red cells is complicated by physicochemical alterations of the ion itself. At concentrations below 5 x 10x4 M, the rate of uptake bears a linear relationship to the Mn++ concentration. The permeability constant for inward movement of Mn++ is 2.87 ± 0.13 (S.E.) x 10-9 cm./sec. The rate is not influenced by the addition of metabolic substrates such as glucose or adenosine or the metabolic inhibitors iodoacetate or fluoride. Co++, Ca++, and Mg++ do not appear to compete with Mn++ for entry, but at high concentrations relative to Mn++, they reduce the rate of entry. Ca++ is far more effective than Co++ or Mg++ in this regard. The permeability constant for outward Mn++ movement is 1.38 ± 0.21 (S.E.) x 10-9 cm./sec., about half of that for entry. This slower rate of outward movement is consistent with the finding that 40 to 60 per cent of the Mn++ taken up by the red cells is non-ultrafilterable. Less than 5 to 10 per cent of the Mn++ appears to be bound to the stroma. It is concluded that entry and exit of Mn++ is a process of passive diffusion involving no carriers, transport, or metabolic linkage.

Protein 4.2 is critical to CD47-membrane skeleton attachment in human red cells

Blood ◽  
2004 ◽  
Vol 103 (3) ◽  
pp. 1131-1136 ◽  
Author(s):  
Kris Noel Dahl ◽  
Ranganath Parthasarathy ◽  
Connie M. Westhoff ◽  
D. Mark Layton ◽  
Dennis E. Discher
Keyword(s):  
Membrane Protein ◽  
Blood Cells ◽  
Significant Variation ◽  
Integral Membrane Protein ◽  
Membrane Skeleton ◽  
Band 3 ◽  
Normal Cells ◽  
Human Red Blood Cells ◽  
Protein 4.2 ◽  
Human Red Cells

Abstract The reduction in expression of the integral membrane protein CD47 in human red blood cells (RBCs) deficient in protein 4.2 suggests that protein 4.2 may mediate a linkage of CD47 to the membrane skeleton. We compared the fractions of membrane skeleton-attached CD47, Rh-associated glycoprotein (RhAG), Rh, and band 3 in normal and protein 4.2-deficient cells using fluorescence-imaged microdeformation. We found that CD47 attachment decreases from 55% in normal cells to 25% to 35% in 4.2-deficient cells. RhAG, which has been shown to have no significant variation in expression among the cells studied, shows a significant decrease in membrane skeleton attachment in 4.2-deficient cells from 60% to 40%. Both Rh and band 3, which have also been shown to have no change in expression, show a smaller decrease from 75% attached in normal RBCs to 55% attached in 4.2-deficient cells. In normal cells, Rh phenotype influences CD47 expression but not the level of membrane skeleton attachment of CD47. In contrast, the results indicate that protein 4.2 strongly influences CD47 levels as well as the extent of membrane skeleton attachment in the RBC, whereas protein 4.2 affects membrane skeletal attachment of RhAG, Rh, and band 3 to a lesser extent. (Blood. 2004;103:1131-1136)

scholarly journals Altered response of stored red cells to Ca2+ stress

Blood ◽  
1985 ◽  
Vol 65 (4) ◽  
pp. 1025-1027 ◽  
Author(s):  
L Lorand ◽  
M Michalska
Keyword(s):  
Membrane Protein ◽  
Band 3 ◽  
Blood Bank ◽  
Red Cells ◽  
Cross Linking ◽  
Short Term ◽  
Important Sign ◽  
Altered Response ◽  
Stored Blood ◽  
In Cells

Short-term Ca2+ loading of erythrocytes was used as a test for probing membrane protein susceptibilities toward intrinsic enzymes in cells from fresh and from stored blood. The proteolytic response, affecting mainly glycophorin and band 3, could be elicited only in fresh cells, whereas the transglutaminase-mediated cross-linking reaction was evident both in fresh and stored cells. Loss of the proteolytic response might be an important sign of erythrocyte damage from blood bank storage.

Membrane lipid diffusion and band 3 protein changes in human erythrocytes due to acute hypobaric hypoxia

1998 ◽  
Vol 275 (6) ◽  
pp. C1429-C1431 ◽  
Author(s):  
Gloria Celedón ◽  
Gustavo González ◽  
Carlos P. Sotomayor ◽  
Claus Behn
Keyword(s):  
Erythrocyte Membrane ◽  
Hypobaric Hypoxia ◽  
Protein Modification ◽  
Membrane Lipids ◽  
Band 3 ◽  
Membrane Lipid ◽  
Erythrocyte Membranes ◽  
Dodecanoic Acid ◽  
Band 3 Protein ◽  
Acute Hypobaric Hypoxia

Because it has been reported that hypoxia in rats may promote lipid peroxidation and other free radical reactions that could modify membrane lipids and proteins, the effect of acute hypobaric hypoxia on human erythrocyte membranes was investigated. 12-(1-Pyrene)dodecanoic acid fluorescent probe was used to assess short-range lateral diffusion status in the membrane bilayer. Membrane protein modification was detected by SDS-PAGE. Healthy young men were exposed for 20 min to the hypobaric hypoxia, simulating an altitude of 4,500 m. Under this condition, erythrocyte membrane lipids reached a state of higher lateral diffusivity with respect to normobaric conditions and membrane band 3 protein was modified, becoming more susceptible to membrane-bound proteinases. These observations suggest that acute hypobaric hypoxia may promote an oxidative stress condition in the erythrocyte membrane.

scholarly journals Altered response of stored red cells to Ca2+ stress

Blood ◽  
1985 ◽  
Vol 65 (4) ◽  
pp. 1025-1027 ◽  
Author(s):  
L Lorand ◽  
M Michalska
Keyword(s):  
Membrane Protein ◽  
Band 3 ◽  
Blood Bank ◽  
Red Cells ◽  
Cross Linking ◽  
Short Term ◽  
Important Sign ◽  
Altered Response ◽  
Stored Blood ◽  
In Cells

Abstract Short-term Ca2+ loading of erythrocytes was used as a test for probing membrane protein susceptibilities toward intrinsic enzymes in cells from fresh and from stored blood. The proteolytic response, affecting mainly glycophorin and band 3, could be elicited only in fresh cells, whereas the transglutaminase-mediated cross-linking reaction was evident both in fresh and stored cells. Loss of the proteolytic response might be an important sign of erythrocyte damage from blood bank storage.

scholarly journals Kinetics and stoichiometry of Na-dependent Li transport in human red blood cells.

1978 ◽  
Vol 72 (2) ◽  
pp. 249-265 ◽  
Author(s):  
B Sarkadi ◽  
J K Alifimoff ◽  
R B Gunn ◽  
D C Tosteson
Keyword(s):  
Transport System ◽  
Red Cells ◽  
Normal Male ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Human Red Blood Cells ◽  
Na Efflux ◽  
Tightly Coupled ◽  
Male Subjects ◽  
Human Red Cells

This paper describes the kinetics and stoichiometry of a tightly coupled Na-Li exchange transport system in human red cells. The system is inhibited by phloretin and furosemide but not by ouabain. Li influx by this system increases and saturates with increasing concentrations of external Li and internal Na and is inhibited competitively by external Na. Comparable functions relate Li efflux and Na efflux to internal and external Li and Na concentrations. Analysis of these relations yields the following values for the ion concentrations required to half-maximally activate the transport system: internal Na and Li 9.0 and 0.5 mM, respectively, external Na and Li 25 and 1.5 mM, respectively. The system performs a 1:1 exchange of Na and Li moving in opposite directions across the red cell membrane. We found no evidence for a simultaneous transport of more than one Na and Li by the system. The maximum transport rate of Na-dependent Li transport varied between 0.1 and 0.37 mmol/(liter of cells X h) in the red cells of the five normal male subjects studied. No significant variations between individual subjects were observed for bicarbonate-stimulated Li transport and for the residual Li fluxes which occur in the absence of bicarbonate and in the presence of ouabain plus phloretin.

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