scholarly journals Control of oxygen affinity of hemoglobin in K562 cells induced by hemin

Blood ◽  
1984 ◽  
Vol 63 (6) ◽  
pp. 1447-1452
Author(s):  
Y Wu ◽  
A Dean ◽  
W Egan ◽  
AN Schechter
Keyword(s):  
Time Course ◽  
31P Nmr ◽  
Oxygen Affinity ◽  
K562 Cells ◽  
Hill Coefficient ◽  
Oxygen Dissociation ◽  
Oxygen Affinity Of Hemoglobin ◽  
Dissociation Curves ◽  
The Relationship ◽  
2,3 Dpg

The oxygen affinity of hemoglobin in K562 cells induced by hemin and the relationship between levels of 2,3-diphosphoglycerate (2,3-DPG) and hemoglobin have been investigated. Absorption spectra of induced cells indicate that the hemoglobin is oxygenated; oxygen dissociation curves are symmetric, with a P50 of 20 +/- 0.9 mm Hg, Hill coefficient of 2.5, and a normal temperature dependence. The intracellular pH measured by phosphorus 31 nuclear magnetic resonance (NMR) was 7.3. The amount of 2,3-DPG was determined by an enzymatic method and by 31P NMR. The level of 2,3-DPG in uninduced K562 cells, containing 0.5 pg of hemoglobin per cell, was low (5 +/- 0.5 mumole/10(8) cells), but increased to 64 +/- 5 mumole/10(8) cells upon induction of hemoglobin accumulation (to a final level of 20 pg hemoglobin/cell). For several experiments, there was a closely coordinated relationship between 2,3-DPG and hemoglobin levels, at about 1:1 stoichiometry of the two molecules. The time course of induction of hemoglobin, and of 2,3-DPG levels, are very similar; both processes are reversible. These data suggest that induction of hemoglobin synthesis in K562 cells by hemin results in hemoglobin-containing cells with normal oxygenation properties and that 2,3-DPG and hemoglobin levels are coordinately controlled in these cells. Elucidation of the mechanism of this effect should be of importance in understanding the erythroid-like differentiation of these cells.

scholarly journals Control of oxygen affinity of hemoglobin in K562 cells induced by hemin

Blood ◽  
1984 ◽  
Vol 63 (6) ◽  
pp. 1447-1452 ◽  
Author(s):  
Y Wu ◽  
A Dean ◽  
W Egan ◽  
AN Schechter
Keyword(s):  
Time Course ◽  
31P Nmr ◽  
Oxygen Affinity ◽  
K562 Cells ◽  
Hill Coefficient ◽  
Oxygen Dissociation ◽  
Oxygen Affinity Of Hemoglobin ◽  
Dissociation Curves ◽  
The Relationship ◽  
2,3 Dpg

Abstract The oxygen affinity of hemoglobin in K562 cells induced by hemin and the relationship between levels of 2,3-diphosphoglycerate (2,3-DPG) and hemoglobin have been investigated. Absorption spectra of induced cells indicate that the hemoglobin is oxygenated; oxygen dissociation curves are symmetric, with a P50 of 20 +/- 0.9 mm Hg, Hill coefficient of 2.5, and a normal temperature dependence. The intracellular pH measured by phosphorus 31 nuclear magnetic resonance (NMR) was 7.3. The amount of 2,3-DPG was determined by an enzymatic method and by 31P NMR. The level of 2,3-DPG in uninduced K562 cells, containing 0.5 pg of hemoglobin per cell, was low (5 +/- 0.5 mumole/10(8) cells), but increased to 64 +/- 5 mumole/10(8) cells upon induction of hemoglobin accumulation (to a final level of 20 pg hemoglobin/cell). For several experiments, there was a closely coordinated relationship between 2,3-DPG and hemoglobin levels, at about 1:1 stoichiometry of the two molecules. The time course of induction of hemoglobin, and of 2,3-DPG levels, are very similar; both processes are reversible. These data suggest that induction of hemoglobin synthesis in K562 cells by hemin results in hemoglobin-containing cells with normal oxygenation properties and that 2,3-DPG and hemoglobin levels are coordinately controlled in these cells. Elucidation of the mechanism of this effect should be of importance in understanding the erythroid-like differentiation of these cells.

Effect of carbon monoxide on equilibrium between oxygen and hemoglobin

1976 ◽  
Vol 230 (2) ◽  
pp. 471-475 ◽  
Author(s):  
Y Okada ◽  
I Tyuma ◽  
Y Ueda ◽  
T Sugimoto
Keyword(s):  
Carbon Monoxide ◽  
Oxygen Saturation ◽  
Whole Blood ◽  
Oxygen Affinity ◽  
Hill Coefficient ◽  
Human Whole Blood ◽  
Oxygen Dissociation ◽  
Dissociation Curves ◽  
The Hill ◽  
Good Agreement

Oxygen dissociation curves of partially CO-saturated human whole blood drawn freshly or preserved more than 3 wk were studied. With increasing CO-hemoglobin concentrations, oxygen affinity of the blood increased and the Hill coefficient, n, fell and gradually approached unity. The changes induced by CO-hemoglobin showed practically no difference in the presence or absence of 2,3-diphosphoglycerate. The Bohr coefficient, deltalog P50/deltapH, was determined as a function of oxygen saturation for various concentrations of CO-hemoglobin. The coefficient remained essentially unchanged in the presence of CO-hemoglobin. In the presence of less than 50% CO-hemoglobin, a good agreement was observed between the observed oxygen dissociation curves and the curves calculated according to Roughton and Darling (Am. J. Physiol. 141: 17-31, 1944). Based on these results, physiological implications of carboxyhemoglobinemia are discussed quantitatively in comparison with methemoglobinemia.

The oxygen dissociation curve of haemoglobin in dilute solution

1936 ◽  
Vol 120 (819) ◽  
pp. 484-495 ◽  
Keyword(s):  
Spectroscopic Method ◽  
Marked Change ◽  
Maternal Blood ◽  
Gas Analysis ◽  
Dilute Solution ◽  
Dissociation Curve ◽  
Foetal Haemoglobin ◽  
Oxygen Dissociation ◽  
Dissociation Curves ◽  
The Relationship

The spectroscopic determination of the oxygen dissociation curves of haemoglobin has an advantage over the tonometer and gas analysis method, in that much smaller quantities of haemoglobin can be made use of. The spectroscopic method was used to determine the relationship between the foetal and maternal haemoglobins in the sheep during a study of foetal respiration made by Barcroft (1935). The conditions for the comparison of the haemoglobins were a dilute solution of the haemoglobin at p H 9·2 (borate buffer) and at 20° C. These conditions were chosen because of the very accurate determinations of the dissociation curves of dilute haemoglobin of the sheep by Forbes and Roughton (1931) and because these authors recommend p H 9·2 at room temperature as most suitable for a study of the oxygen equilibrium of haemoglobin, all the haemoglobin being in the form of the alkali salt. McCarthy (1933) and Hall (1934) had found previously that the haemoglobins of the foetal and maternal goat were different, the foetal haemoglobin (in the blood and as purified haemoglobin) having a higher affinity for oxygen. The same relationship was found to exist in the sheep haemoglobins in dilute solution at 20° C and p H 9·2. When samples of human foetal and maternal blood (sent by Professor Fleming from the Obstetrical Department of the Royal Free Hospital) were compared in dilute solution it was found that the foetal haemoglobin had a lower affinity for oxygen than the maternal. This was also found by Haurowitz (1935) for dilute solutions of the haemoglobins of mother and new born infant. Haurowitz, however, pointed out that in the corpuscles the affinity for oxygen is less in the infant’s haemoglobin than in that of the mother, but the method used by him did not allow of measurements on suspensions of corpuscles. In the present work the dissociation curves of dilute suspensions of corpuscles have been compared with similar solutions of the haemoglobin. It was found that the relationship of the dissociation curves for human foetal and maternal corpuscles is the same as that found by Barcroft in the goat and in the sheep. It has now been found that by a dilution of human adult haemoglobin the dissociation curve is altered by 200% to a position of higher affinity for oxygen, without any marked change in shape. The haemoglobin of the human foetus, on the other hand, is much less affected by dilution, thus explaining the anomaly of the reversed relationship when solutions of the haemoglobins are used instead of suspensions of corpuscles. It was shown by the work of Bock, Field, and Adair (1924), and by Adair (1925), that a solution of haemoglobin free from stromata and of a similar concentration to blood gives a dissociation curve like whole blood. This makes it clear that in the comparison of dilute haemoglobin solutions with suspensions of corpuscles we are concerned, not simply with a change in the haemoglobin due to haemolysis, but a change due to a dilution of the contents of the corpuscle.

An electrolytic method for determining oxygen dissociation curves using small blood samples: the effect of temperature on trout and human blood

1976 ◽  
Vol 65 (1) ◽  
pp. 21-38
Author(s):  
G. M. Hughes ◽  
J. G. O'Neill ◽  
W.J. van Aardt
Keyword(s):  
Human Blood ◽  
Marked Effect ◽  
Effect Of Temperature ◽  
Oxygen Dissociation Curve ◽  
Dissociation Curve ◽  
Oxygen Dissociation ◽  
Fish Blood ◽  
Bohr Factor ◽  
Dissociation Curves ◽  
The Relationship

1. A detailed account is given of an electrolytic method for determining the oxygen dissociation curve of fish blood using a single sample of 50–100 mul for the whole curve. The accuracy and some of the problems arising from its uses are discussed. 2. Oxygen dissociation curves have been determined for trout blood and human blood at temperatures of 15 and 37 degrees C. The relationship between P50 and temperature is similar to that obtained using other methods. Absolute values of P50 are generally lower than those obtained by other methods, especially in the case of fish blood. 3. The effect of PCO2 and pH on the oxygen dissociation curve of trout blood is tested and it is shown that PCO2 has a more marked effect than pH when the other factor is maintained at a constant level. The Bohr factor (delta log P50/delta pH) appears to be approximately the same and independent of the PCO2. 4.The P50 of ray blood determined from fish during and after an operation showed an increased Bohr factor.

Postnatal regulation of canine oxygen delivery: erythrocyte components affecting Hb function

1980 ◽  
Vol 238 (1) ◽  
pp. H73-H79 ◽  
Author(s):  
P. A. Mueggler ◽  
G. Jones ◽  
J. S. Peterson ◽  
J. M. Bissonnette ◽  
R. D. Koler ◽  
...  
Keyword(s):  
Oxygen Affinity ◽  
Fetal Hemoglobin ◽  
Blood Oxygen ◽  
Oxygen Dissociation ◽  
Affinity Change ◽  
Blood Oxygen Affinity ◽  
Postnatal Change ◽  
2,3 Dpg

A rightward shift in the blood oxygen dissociation curve occurs during the 1st mo of canine life. A detailed peptide analysis indicated that dogs do not have a separate fetal hemoglobin. Other erythrocyte components such as ATP, K+, Na+, and H+ were excluded as significant mediators of the postnatal oxygen affinity change. Erythrocyte 2,3-DPG levels essentially zero in fetal dogs, increased rapidly during the 1st mo of canine life. There was a significant correlation between this postnatal 2,3-DPG increase and the postnatal decrease in blood oxygen affinity. Dialyzed hemolysates of fetal or adult canine blood have the same intrinsic oxygen affinity and the same response to normal adult levels of 2,3-DPG. Furthermore, the magnitude and direction of this 2,3-DPG-induced decrease in oxygen affinity in vitro are comparable to the in vivo postnatal change in oxygen affinity.

Conventional Haemodialysis and Ultrafiltration: Effects on Erythrocyte 2,3-Diphosphoglycerate and Oxygen Affinity in Patients with Uraemia

1980 ◽  
Vol 59 (2) ◽  
pp. 143-145 ◽  
Author(s):  
G. Severini ◽  
A. Giuliani ◽  
D. Maffi ◽  
N. Cerulli
Keyword(s):  
Oxygen Affinity ◽  
Blood Ph ◽  
The Relationship ◽  
2,3 Dpg

1. The relationship between erythrocyte 2,3-diphosphoglycerate (2,3-DPG) and changes in blood pH and in oxygen affinity were studied in six patients treated with two dialysis techniques: conventional haemodialysis and ultrafiltration followed by conventional haemodialysis. 2. The decrease in erythrocyte 2,3-diphosphoglycerate and increase in pH after conventional haemodialysis may significantly increase the affinity of haemoglobin for oxygen and consequently result in inadequate oxygenation of tissue. 3. Ultrafiltration followed by conventional haemodialysis, on the contrary, is more beneficial for oxygenation of tissue and post-dialysis symptoms may be reduced.

scholarly journals Treatment of sickle cell disease by increasing oxygen affinity of hemoglobin

Blood ◽  
2021 ◽  
Author(s):  
Eric R Henry ◽  
Belhu Metaferia ◽  
Quan Li ◽  
Julia Harper ◽  
Robert B Best ◽  
...  
Keyword(s):  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Oxygen Delivery ◽  
Cell Damage ◽  
Oxygen Affinity ◽  
Red Cell ◽  
Cell Disease ◽  
Oxygen Dissociation ◽  
Dissociation Curves

The issue of treating sickle cell disease with drugs that increase hemoglobin oxygen affinity has come to the fore with the FDA approval in 2019 of voxelotor, the only anti-sickling drug approved since hydroxyurea in 1998. Voxelotor reduces sickling by increasing the concentration of the non-polymerizing, high oxygen affinity R (oxy) conformation of HbS. Treatment of sickle cell patients with voxelotor increases Hb levels and decreases indicators of hemolysis, but with no indication as yet that it reduces the frequency of pain episodes. Here we use the allosteric model of Monod, Wyman, and Changeux to simulate whole blood oxygen dissociation curves and red cell sickling in the absence and presence of voxelotor under the in vivo conditions of rapid oxygen pressure decreases. Our modeling agrees with experiments using a new robust assay, which shows the very large, expected decrease in sickling from the drug. The modeling indicates, however, that the increase in oxygen delivery from reduced sickling is largely offset by the increase in oxygen affinity. The net result is that the drug increases overall oxygen delivery only at the very lowest oxygen pressures. Reduction of sickling does, however, mitigate against red cell damage and explains the observed decrease in hemolysis. More importantly, our modeling of in vivo oxygen dissociation, sickling, and oxygen delivery suggests that drugs that increase fetal hemoglobin or decrease MCHC, should be more therapeutically effective than drugs that increase oxygen affinity.

Oxygen dissociation curve for chorioallantoic capillary blood of chicken embryo

1976 ◽  
Vol 40 (3) ◽  
pp. 393-398 ◽  
Author(s):  
H. Tazawa ◽  
T. Ono ◽  
M. Mochizuki
Keyword(s):  
Oxygen Affinity ◽  
Capillary Blood ◽  
Oxygen Dissociation Curve ◽  
Dissociation Curve ◽  
Oxygen Dissociation ◽  
Oxygen Capacity ◽  
Bohr Factor ◽  
Hill’S Equations ◽  
Dissociation Curves ◽  
Clear Relation

Oxygen dissociation curves for blood in the chorioallantoic capillary of chicken embryos were determined using a microphotometric apparatus made for measuring the reaction velocity of a red blood cell with oxygen and carbon monoxide. The modified Hill's equations expressing the dissociation curve during development were calculated by two methods. P50's at pH of 7.4 were found to be 60.0, 54.4, 46.2, 33.1, and 28.6 mmHg for 10, 12, 14, 16 and 18 days of incubation, respectively. Although the Bohr factor did not show a clear relation to age, the oxygen affinity and the oxygen capacity tended to increase with the lapse of days, and the power of heme-to-heme interaction, to decrease with age. The findings imply that there is a respiratory adaptation of embryos during development.

scholarly journals Dose- and Sex-Dependent Changes in Hemoglobin Oxygen Affinity by the Micronutrient 5-Hydroxymethylfurfural and α-Ketoglutaric Acid

Nutrients ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 3448
Author(s):  
Simon Woyke ◽  
Norbert Mair ◽  
Astrid Ortner ◽  
Thomas Haller ◽  
Marco Ronzani ◽  
...  
Keyword(s):  
Linear Regression Analysis ◽  
Oxygen Affinity ◽  
Nutritional Supplement ◽  
Hill Coefficient ◽  
Therapeutic Effects ◽  
Oxygen Dissociation ◽  
Hemoglobin Oxygen Affinity ◽  
Clinical Conditions

5-Hydroxymethylfurfural (5-HMF) is known to increase hemoglobin oxygen affinity (Hb–O2 affinity) and to induce a left shift of the oxygen dissociation curve (ODC). It is under investigation as a therapeutic agent in sickle cell anemia and in conditions where pulmonary oxygen uptake is deteriorated or limited (e.g., various clinical conditions or altitude exposure). The combination of 5-HMF and α-ketoglutaric acid (αKG) is commercially available as a nutritional supplement. To further elucidate dose effects, ODCs were measured in vitro in venous whole blood samples of 20 healthy volunteers (10 female and 10 male) after the addition of three different doses of 5-HMF, αKG and the combination of both. Linear regression analysis revealed a strong dose-dependent increase in Hb–O2 affinity for 5-HMF (R2 = 0.887; p < 0.001) and the commercially available combination with αKG (R2 = 0.882; p < 0.001). αKG alone increased Hb–O2 affinity as well but to a lower extent. Both the combination (5-HMF + αKG) and 5-HMF alone exerted different P50 and Hill coefficient responses overall and between sexes, with more pronounced effects in females. With increasing Hb–O2 affinity, the sigmoidal shape of the ODC was better preserved by the combination of 5-HMF and αKG than by 5-HMF alone. Concerning the therapeutic effects of 5-HMF, this study emphasizes the importance of adequate dosing in various physiological and clinical conditions, where a left-shifted ODC might be beneficial. By preserving the sigmoidal shape of the ODC, the combination of 5-HMF and αKG at low (both sexes) and medium (males only) doses might be able to better maintain efficient oxygen transport, particularly by mitigating potentially deteriorated oxygen unloading in the tissue. However, expanding knowledge on the interaction between 5-HMF and Hb–O2 affinity in vitro necessitates further investigations in vivo to additionally assess pharmacokinetic mechanisms.

The Lack of Effects of Phototherapy on Neonatal Oxygen Dissociation Curves and Hemoglobin Concentration in Vivo

PEDIATRICS ◽  
1977 ◽  
Vol 59 (6) ◽  
pp. 1027-1031
Author(s):  
Barry D. Chandler ◽  
William J. Cashore ◽  
Pierre J. P. Monin ◽  
William Oh
Keyword(s):  
Serum Bilirubin ◽  
Hemoglobin Concentration ◽  
Control Group ◽  
Dissociation Curve ◽  
Oxygen Dissociation ◽  
The Right ◽  
Dissociation Curves ◽  
2,3 Dpg

Phototherapy has been shown to cause hemolysis of fetal red cells and a shift to the right in the neonatal oxygen dissociation curve (ΔP50) in vitro. To determine if these parameters act similarly in vivo, we have studied 16 icteric infants before and after phototherapy and compared them with eight control infants studied at birth and at 3 days of age, measuring the change in P50, 2,3-diphosphoglycerate (2,3-DPG), serum bilirubin, the percent of bilirubin/albumin saturation, and the hemoglobin concentration in the two groups. Following phototherapy, in the icteric infants there was a shift to the right in the O2 dissociation curve of + 1.7 mm Hg, a rise in 2,3-DPG of 2.26 µmol/gm Hb, a fall in serum bilirubin of 4.3 mg/100 ml, a decrease in percent bilirubin/albumin saturation of 12.4%, and a drop in hemoglobin of 1.1 gm/100 ml. The control group showed a ΔP50 of + 2.0 mm Hg, a rise in 2,3-DPG of 3.67 µmol/gm Hb, an increase in serum bilirubin of 3.2 mg/100 ml, an increase in percent bilirubin/albumin saturation of 9.3%, and a fall in hemoglobin of 0.3 gm/100 ml. Significant differences between the groups were seen only in the changes in bilirubin concentration and percent bilirubin/albunmin saturation. The magnitude of changes in P50, 2,3-DPG, and hemoglobin concentration was similar in the phototherapy and control groups and was related to the expected changes with reference to postnatal age. These results suggest that phototherapy in vivo neither affects fetal erythrocytic affinity for oxygen nor causes hemolysis.

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