Bohr effect

Not to be confused with theBohr equation.

TheBohr effectis a phenomenon first described in 1904 by the Danish physiologistChristian Bohr.Hemoglobin's oxygen binding affinity (seeoxygen–haemoglobin dissociation curve) is inversely related both to acidity and to the concentration of carbon dioxide.That is, the Bohr effect refers to the shift in the oxygen dissociation curve caused by changes in the concentration ofcarbon dioxideor thepHof the environment. Since carbon dioxide reacts with water to formcarbonic acid,an increase in CO₂results in a decrease in bloodpH,resulting in hemoglobin proteins releasing their load of oxygen. Conversely, a decrease in carbon dioxide provokes an increase in pH, which results in hemoglobin picking up more oxygen.

Christian Bohr, who was credited with the discovery of the effect in 1904.

Experimental discovery

The original dissociation curves from Bohr's experiments in the first description of the Bohr effect, showing a decrease in oxygen affinity as the partial pressure of carbon dioxide increases. This is also one of the first examples ofcooperative binding.X-axis: oxygen partial pressure inmmHg,Y-axis %oxy-hemoglobin.The curves were obtained using wholedogblood,with the exception of the dashed curve, for whichhorsebloodwas used.

In the early 1900s, Christian Bohr was a professor at theUniversity of Copenhagenin Denmark, already well known for his work in the field of respiratory physiology.He had spent the last two decades studying the solubility of oxygen, carbon dioxide, and other gases in various liquids,and had conducted extensive research on haemoglobin and its affinity for oxygen.In 1903, he began working closely withKarl HasselbalchandAugust Krogh,two of his associates at the university, in an attempt to experimentally replicate the work ofGustav von Hüfner,using whole blood instead of haemoglobin solution.Hüfner had suggested that the oxygen-haemoglobin binding curve washyperbolicin shape,but after extensive experimentation, the Copenhagen group determined that the curve was in factsigmoidal.Furthermore, in the process of plotting out numerous dissociation curves, it soon became apparent that high partial pressures of carbon dioxide caused the curves to shift to the right.Further experimentation while varying the CO₂concentration quickly provided conclusive evidence, confirming the existence of what would soon become known as the Bohr effect.

Controversy

There is some more debate over whether Bohr was actually the first to discover the relationship between CO₂and oxygen affinity, or whether the Russian physiologistBronislav Verigobeat him to it, allegedly discovering the effect in 1898, six years before Bohr.While this has never been proven, Verigo did in fact publish a paper on the haemoglobin-CO₂relationship in 1892.His proposed model was flawed, and Bohr harshly criticized it in his own publications.

Another challenge to Bohr's discovery comes from within his lab. Though Bohr was quick to take full credit, his associate Krogh, who invented the apparatus used to measure gas concentrations in the experiments,maintained throughout his life that he himself had actually been the first to demonstrate the effect. Though there is some evidence to support this, retroactively changing the name of a well-known phenomenon would be extremely impractical, so it remains known as the Bohr effect.

Physiological role

The Bohr effect increases the efficiency of oxygen transportation through the blood. After hemoglobin binds to oxygen in thelungsdue to the high oxygen concentrations, the Bohr effect facilitates its release in the tissues, particularly those tissues in most need of oxygen. When a tissue's metabolic rate increases, so does its carbon dioxide waste production. When released into the bloodstream, carbon dioxide formsbicarbonateand protons through the following reaction:

${\ce {CO2 + H2O <=> H2CO3 <=> H+ + HCO3^-}}$

Although this reaction usually proceeds very slowly, the enzymecarbonic anhydrase(which is present inred blood cells) drastically speeds up the conversion to bicarbonate and protons.This causes the pH of the blood to decrease, which promotes the dissociation of oxygen from haemoglobin, and allows the surrounding tissues to obtain enough oxygen to meet their demands. In areas where oxygen concentration is high, such as the lungs, binding of oxygen causes haemoglobin to release protons, which recombine with bicarbonate to eliminate carbon dioxide duringexhalation.These opposing protonation and deprotonation reactions occur in equilibrium resulting in little overall change in blood pH.

The Bohr effect enables the body to adapt to changing conditions and makes it possible to supply extra oxygen to tissues that need it the most. For example, whenmusclesare undergoing strenuous activity, they require large amounts of oxygen to conductcellular respiration,which generates CO₂(and therefore HCO₃⁻and H⁺) as byproducts. These waste products lower the pH of the blood, which increases oxygen delivery to the active muscles. Carbon dioxide is not the only molecule that can trigger the Bohr effect. If muscle cells aren't receiving enough oxygen for cellular respiration, they resort tolactic acid fermentation,which releaseslactic acidas a byproduct. This increases the acidity of the blood far more than CO₂alone, which reflects the cells' even greater need for oxygen. In fact, under anaerobic conditions, muscles generate lactic acid so quickly that pH of the blood passing through themuscleswill drop to around 7.2, which causes haemoglobin to begin releasing roughly 10% more oxygen.

The magnitude of the Bohr effect is given by${\textstyle {\scriptstyle \Delta \log(P_{50}) \over \Delta {\text{pH}}}}$,which is the slope on this graph. A steeper slope means a stronger Bohr effect.

Strength of the effect and body size

The magnitude of the Bohr effect is usually given by the slope of the${\textstyle \log(P_{50})}$vs${\textstyle {\text{pH}}}$curve where,P₅₀refers to the partial pressure of oxygen when 50% of haemoglobin's binding sites are occupied. The slope is denoted:${\textstyle {\scriptstyle \Delta \log(P_{50}) \over \Delta {\text{pH}}}}$where${\textstyle \Delta }$denotes change. That is,${\textstyle \Delta \log(P_{50})}$denotes the change in${\textstyle \log(P_{50})}$and${\textstyle \Delta {\text{pH}}}$the change in${\textstyle {\text{pH}}}$. Bohr effect strength exhibits an inverse relationship with the size of an organism: the magnitude increases as size and weight decreases. For example,micepossess a very strong Bohr effect, with a${\textstyle {\scriptstyle \Delta \log(P_{50}) \over \Delta {\text{pH}}}}$value of -0.96, which requires relatively minor changes in H⁺or CO₂concentrations, whileelephantsrequire much larger changes in concentration to achieve a much weaker effect${\textstyle \left({\scriptstyle \Delta \log(P_{50}) \over \Delta {\text{pH}}}=-0.38\right)}$.

Mechanism

Allosteric interactions

Haemoglobin changes conformation from a high-affinity R state (oxygenated) to a low-affinity T state (deoxygenated) to improve oxygen uptake and delivery.

The Bohr effect hinges around allosteric interactions between thehemesof the haemoglobintetramer,a mechanism first proposed by Max Perutz in 1970.Haemoglobin exists in two conformations: a high-affinity R state and a low-affinity T state. When oxygen concentration levels are high, as in the lungs, the R state is favored, enabling the maximum amount of oxygen to be bound to the hemes. In the capillaries, where oxygen concentration levels are lower, the T state is favored, in order to facilitate the delivery of oxygen to the tissues. The Bohr effect is dependent on this allostery, as increases in CO₂and H⁺help stabilize the T state and ensure greater oxygen delivery to muscles during periods of elevated cellular respiration. This is evidenced by the fact thatmyoglobin,amonomerwith no allostery, does not exhibit the Bohr effect.Haemoglobin mutants with weaker allostery may exhibit a reduced Bohr effect. For example, in Hiroshima varianthaemoglobinopathy,allostery in haemoglobin is reduced, and the Bohr effect is diminished. As a result, during periods of exercise, the mutant haemoglobin has a higher affinity for oxygen and tissue may suffer minoroxygen starvation.

T-state stabilization

When hemoglobin is in its T state, theN-terminalamino groups of the α-subunits and theC-terminalhistidineof the β-subunits are protonated, giving them a positive charge and allowing these residues to participate inionic interactionswith carboxyl groups on nearby residues. These interactions help hold the haemoglobin in the T state. Decreases in pH (increases in acidity) stabilize this state even more, since a decrease in pH makes these residues even more likely to be protonated, strengthening the ionic interactions. In the R state, the ionic pairings are absent, meaning that the R state's stability increases when the pH increases, as these residues are less likely to stay protonated in a more basic environment. The Bohr effect works by simultaneously destabilizing the high-affinity R state and stabilizing the low-affinity T state, which leads to an overall decrease in oxygen affinity.This can be visualized on anoxygen-haemoglobin dissociation curveby shifting the whole curve to the right.

Carbon dioxide can also react directly with the N-terminal amino groups to formcarbamates,according to the following reaction:

${\ce {R-NH2 + CO2 <=> R-NH-COO^- + H+}}$

CO₂forms carbamates more frequently with the T state, which helps to stabilize this conformation. The process also creates protons, meaning that the formation of carbamates also contributes to the strengthening of ionic interactions, further stabilizing the T state.

Special cases

Though they are one of the largest animals on the planet, humpback whales have a Bohr effect magnitude similar to that of a guinea pig.

Marine mammals

An exception to the otherwise well-supported link between animal body size and the sensitivity of its haemoglobin to changes in pH was discovered in 1961.Based on their size and weight, manymarine mammalswere hypothesized to have a very low, almost negligible Bohr effect.However, when their blood was examined, this was not the case.Humpback whalesweighing 41,000 kilograms had an observed${\textstyle {\scriptstyle \Delta \log(P_{50}) \over \Delta {\text{pH}}}}$value of 0.82, which is roughly equivalent to the Bohr effect magnitude in a 0.57 kgguinea pig.This extremely strong Bohr effect is hypothesized to be one of marine mammals' many adaptations for deep, long dives, as it allows for virtually all of the bound oxygen on haemoglobin to dissociate and supply the whale's body while it is underwater.Examination of other marine mammal species supports this. Inpilot whalesandporpoises,which are primarily surface feeders and seldom dive for more than a few minutes, the${\textstyle {\scriptstyle \Delta \log(P_{50}) \over \Delta {\text{pH}}}}$was 0.52, comparable to acow,which is much closer to the expected Bohr effect magnitude for animals of their size.

Carbon monoxide

Another special case of the Bohr effect occurs whencarbon monoxideis present. This molecule serves as acompetitive inhibitorfor oxygen, and binds to haemoglobin to formcarboxyhaemoglobin.Haemoglobin's affinity for CO is about 210 times stronger than its affinity for O₂,meaning that it is very unlikely to dissociate, and once bound, it blocks the binding of O₂to that subunit. At the same time, CO is structurally similar enough to O₂to cause carboxyhemoglobin to favor the R state, raising the oxygen affinity of the remaining unoccupied subunits. This combination significantly reduces the delivery of oxygen to the tissues of the body, which is what makes carbon monoxide sotoxic.This toxicity is reduced slightly by an increase in the strength of the Bohr effect in the presence of carboxyhemoglobin. This increase is ultimately due to differences in interactions between heme groups in carboxyhemoglobin relative to oxygenated hemoglobin. It is most pronounced when the oxygen concentration is extremely low, as a last-ditch effort when the need for oxygen delivery becomes critical. However, the physiological implications of this phenomenon remain unclear.

See also

• Allosteric regulation
• Haldane effect
• Root effect
• Chloride shift

External links

• Impact of training