The Bohr/Haldane effect: a model-based uncovering of the full extent of its impact on O2 delivery to and CO2 removal from tissues

For a century, the influence of the Bohr effect on the utilization of blood-borne oxygen has been deemed secondary to its influence on the uptake of carbon dioxide by the blood. Here, we show that the opposite is the case. Using a simple two-ligand, two-state formulation, we modeled the simultaneous oxygen and proton binding to hemoglobin, as well as the resulting acid-base changes of the surrounding solution. Blocking of the Bohr effect in this model system results in a dramatic increase in the oxygen affinity, as expressed by the oxygen partial pressure at half saturation, the P50. It also becomes clear that the P50 and the Bohr factor (a measure of the size of the Bohr effect) are not independent but directly related. Thus, everything else being equal, varying the number of Bohr groups from 0 to 8 per tetramer results in an increase in the Bohr factor from 0 to −0.9 and an increase in P50 from 6 to 46 mmHg at a constant Pco2 of 40 mmHg. Therefore, changes in hemoglobin structure that lead to changes in the Bohr factor will inevitably also change hemoglobin oxygen affinity. NEW & NOTEWORTHY Using a mathematical model, we show that the Bohr effect has a more profound effect on gas exchange than is evident when comparing oxygen equilibrium curves measured in the laboratory at different constant values of Pco2 or pH. Protons preloaded on the Bohr groups, as well as the protons taken up during oxygen unloading, dramatically decrease oxygen affinity of the physiological oxygen equilibrium curve. Therefore, the Bohr effect is instrumental in setting the oxygen affinity.

Temperature and CO2 Effects on Blood O2 Equilibria in Northern Squawfish, Ptychocheilus oregonensis

In vitro blood O2 equilibrium curves were constructed at 9, 15, 18, and 21 °C from temperature-acclimated northern squawfish, Ptychocheilus oregonensis. At low [Formula: see text] (<1 mm Hg, 1 mm Hg = 133.32 Pa), P50s generally showed variable increases with temperature from 3.6 mm Hg at 9 °C to 8.7 mm Hg at 21 °C, leading to whole-blood temperature effects (ΔH, kilocalories per mole O2) ranging from a low +4.4 at 15–18 °C to a peak −21.2 at 18–21 °C. High-[Formula: see text] (7.6 mm Hg) conditions decreased blood pH and increased P50s at each temperature (Bohr factor). Bohr factors (Φ) ranged from −0.46 at 21 °C to −0.70 at 18 °C. Considered together, ΔH and Φ values suggest an optimal temperature range of 15–18 °C for hemoglobin O2 loading and unloading in northern squawfish. Nonbicarbonate buffer values ranged from −10.04 at 21 °C to −14.13 at 9 °C. Overall, the high O2 affinities and hyperbolic blood O2 equilibrium curves of northern squawfish resemble those of other large cyprinids (e.g., common carp, Cyprinus carpio, tench, Tinca tinca, Sacramento blackfish, Orthodon microlepidotus) indicating a better ability to tolerate hypoxic environments than sympatric rainbow trout, Oncorhynchus mykiss. High northern squawfish blood O2 capacities and Φs suggest high aerobic capacity, especially at temperatures <21 °C.

Blood gas transport and oxygen consumption in a supralittoral crab, Leptograpsus variegatus (Crustacea: Brachyura)

Routine oxygen consumption of Leptograpsus variegatus in air (MO2=6.12 mass (g)0.854 �mol h-1) was elevated from three to six times (depending on body size) following exhausting exercise (MO2= 42.8 mass0.608 �mol h-1). In addition, considerable capacity for anaerobic exercise was apparent and total body L-lactate rose 4.8 times during the exercise period (25-g crab). The respiratory pigment (haemocyanin) had a low affinity under normoxic resting conditions (P50=25 Torr, where 1 Torr = 101 325/760 Pa) but showed a large Bohr effect (Bohr factor = -0.78), a marked temperature sensitivity (ΔH= 67.4 kJ mol-1), and a large potentiating effect by L-lactate (lactate coefficient = - 0.205). These features are more characteristic of the haemocyanin of aquatic crustaceans than those of terrestrial crustaceans. The affinity of the haemocyanin of L. variegatus was also decreased by elevated PCO2. Haemocyanin was responsible for 92% of oxygen transport in both resting and exercised crabs. These features are discussed in relation to the degree of adaptation of L. variegatus to a terrestrial lifestyle.

Human whole-blood O2 affinity: effect of CO2

The proton Bohr factor (phi H = alpha log PO2/alpha pH), the carbamate Bohr factor (phi C = alpha log PO2/alpha log PCO2), the total Bohr factor (phi HC = d log PO2/dpH[base excess) and the CO2 buffer factor (d log PCO2/dpH) were determined in the blood of 12 healthy donors over the whole O2 saturation (SO2) range. All three Bohr factors proved to be dependent on SO2, although to a lesser extent than reported in some of the recent literature. At SO2 = 50% and 37 degrees C, we found phi H = -0.428 +/- 0.010 (SE), phi C = 0.054 +/- 0.006, and phi HC = -0.488 +/- 0.007. The values obtained for phi H, phi C, and d log PCO2/dpH were used to calculate phi HC. Calculated and measured values of phi HC proved to be in good agreement. In an additional series of 12 specimens of human blood we determined the influence of PCO2 on phi H and the influence of pH on phi C. At SO2 = 50%, phi H varied from -0.49 +/- 0.009 at PCO2 = 15 Torr to -0.31 +/- 0.010 at PCO2 = 105 Torr and phi C from 0.157 +/- 0.015 at pH = 7.80 to 0.006 +/- 0.009 at pH = 7.00. When on the basis of these data a second-order term is taken into account, a still slightly better agreement between measured and calculated values of phi HC can be attained.

Oxygen affinity and Bohr coefficients of dog blood

Complete dynamic oxygen equilibrium curves (O2EC) on dog whole blood were measured at 25 and 39 degrees C using a spectrophotometric micro blood film technique. O2EC were run at three CO2 levels (2, 4, and 8%) for each of three base excess levels (-10, 0, +10 meq/l). The standard curve (ph 7.4) was determined for saturations 013;0.98. At 39 degrees C the standard curve O2 pressure at half-saturation (P50) was 31.5 Torr; fixed-acid Bohr factor, -0.488; CO2 Bohr factor, -0.498; delta log P50/delta log PCO2, -0.0045. CO2 Bohr slope was linear over the pH range of 7–8. Bohr factors were not significantly saturation dependent. At 25 degrees C P50 was 15.4 Torr and CO2 Bohr factor, -0.647. The temperature coefficient (delta log P50/delta T) equaled 0.022. Dog O2EC were shown with curve-fitting techniques to be isomorphic with human blood O2EC. The absence of significant oxylabile carbamate formation in dog red blood cells (RBC) was attributed to high 2,3-diphosphoglycerate (DPG) concentrations, 6.23 mM/l RBC, equal to a DPG/Hb4 ratio of 1.12. A simple two-constant equation S = [(37,900)/(P3 + 205P) + 1]-1, where S is saturation and P is oxygen tension, was found to fit the dog 39 degrees C standard curve.

Influence of temperature on hemoglobin-ligand interaction in whole blood

Temperature-dependent change in hemoglobin-oxygen affinity was measured as a function of hemoglobin-oxygen saturation. In addition, the CO2 Bohr factor and fixed acid Bohr factor were measured as a function of saturation of temperatures of 23, 30, 37, and 44 degrees C. Measurements were made on normal blood and blood with reduced 2,3-diphosphoglycerate (DPG). The influence of temperature is greatest at low saturation and is enhanced slightly by DPG depletion. The CO2 Bohr factor is increased at high temperatures; this is primarily due to increased carbamino formation with rising temperature, especially at lower oxygen saturation. The effect of DPG on oxygen affinity is reduced at a high temperature and elevated at low temperature. These diverse effects of temperature on hemoglobin-ligand interaction require consideration in assessing oxygen delivery when temperature is increased or decreased.

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

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.

Oxygen dissociation curve for chorioallantoic capillary blood of chicken embryo

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.