oxygen-hemoglobin dissociation curve

The oxygen-hemoglobin dissociation curve shows how the hemoglobin saturation

with oxygen (SO2,), is related to the partial pressure of oxygen in the blood (PO2).
Hemoglobin is the main protein within red blood cells, and it’s made of four globin
subunits, each containing a heme group capable of binding one molecule of O2.
So each hemoglobin protein can bind 4 molecules of oxygen. But each hemoglobin
isn’t always 100% saturated or bound by oxygen.
A hemoglobin molecule might have no oxygen bound, and be 0% saturated, called
deoxyhemoglobin, and it will take on a tense state shape, or T-state; or it might
have one oxygen bound and three open spots, meaning that particular protein
would be 25% saturated; or two filled spots and two open spots—50%; or 3 spots
filled and one spot open—75%, or all spots filled and 100% saturated.
All of these states - where oxygen is bound to hemoglobin - are called
oxyhemoglobin, changing to its relaxed state, or R-state with each O2 molecule
that binds.
And since there are millions of hemoglobin molecules in a single cell and millions
of red blood cells, the hemoglobin saturation of oxygen is the average saturation
among all of these proteins.
Now it turns out that hemoglobin absorbs different wavelengths of light as it gets
more and more oxygenated.
A technique called pulse oximetry uses this property of hemoglobin to figure out
the average oxygen saturation across millions of hemoglobin proteins.
The main factor that influences oxygen saturation is the partial pressure of oxygen
in the blood, measured in millimeters of mercury (mm Hg).
So for example, at a partial pressure of 25mmHg, hemoglobin proteins might be
50% saturated, called P50; and at a partial pressure of 100mmHg, they might be
98% saturated, meaning most are fully saturated.
And when these points are plotted, the curve takes on a sigmoidal shape.
In practical terms, this sigmoidal shape means that hemoglobin has an increasing
affinity for O2 as the number of bound O2 molecules goes up.
So binding that 4th O2 molecule is much easier than binding that first O2 molecule.
This is called positive cooperativity.
Around 60mmHg, the vast majority of the hemoglobin subunits have bound
oxygen, so the curve starts to level off.
That’s why in arterial blood where the partial pressure of oxygen is around
100mmHg, hemoglobin get fully saturated with oxygen.
And why in the venous capillaries of tissues, where the partial pressure of oxygen
is about 40mmHg, hemoglobin is only about 75% saturated with oxygen.
In other words, about a quarter of the oxygen that’s bound to the hemoglobin gets
dropped off, or unloaded, in the tissues.
Now, there are a few factors that can cause hemoglobin’s affinity for O2 to change.
For example, when CO2 is produced during aerobic metabolism in the tissues, it
dissolves into the blood plasma, increasing the PCO2, and increasing the amount
of CO2 that gets inside the red blood cells.
Inside the red blood cell, the enzyme carbonic anhydrase catalyzes a reaction with
CO2 and water which forms carbonic acid (H2CO3).
Carbonic acid (H2CO3) then splits into a bicarbonate ion (HCO3-) and a hydrogen
H+ ions (H+).
As more and more H+ start to pile up, the pH starts to fall.
Now, both CO2 and H+ can bind to hemoglobin.
CO2 binds to the terminal amino acids in the globin subunits, forming
carbaminohaemoglobin; while H+ bind to amino acid side chains that make up the
globin subunits.
And even though CO2 and H+ don’t compete for the same binding sites as O2 in
hemoglobin, they do stabilize the T-state of hemoglobin.
This results in a decrease in hemoglobin’s affinity for oxygen, forcing the unloading
of O2.
So, for example, when muscles are working hard, like during exercise, they are
producing a lot more CO2, and, as a result, more H+ , which causes the pH to drop.
So O2 is unloaded in the issues that need it most.
Another factor is 2,3-diphosphoglyceric acid (2,3-DPG), which is a metabolic
byproduct of glycolysis in red blood cells.
When there are hypoxic conditions, like a lack of oxygen, which might develop
when a person is at high altitude, 2,3-DPG production increases.
In the middle of the hemoglobin there is a positively charged pocket between the
four globin chains where the negatively charged 2,3-DPG is able to bind.
Again, even though 2,3-DPG doesn’t bind in the same place as O2, it stabilizes the
T-state of hemoglobin and decreases hemoglobin’s affinity for oxygen, forcing the
unloading of O2 in tissues that are in need of it.
Temperature also affects the strength of the bond between oxygen and
hemoglobin.
The heme component of hemoglobin, which is where the oxygen binds, contains
iron, which helps anchor oxygen to hemoglobin.
Higher temperatures affects this bond between iron and oxygen, favoring the T-
state of hemoglobin.
So, using exercise as an example again, increased activity causes muscles to
generate increased heat. And the rise in even a few degrees in temperature allows
for more unloading of O2 , and that helps meet the increased metabolic demands
of the muscles.
Finally, hemoglobin’s affinity for oxygen can depend on the type of hemoglobin.
For example, Hemoglobin A (HbA), which is the main adult hemoglobin has a lower
affinity for oxygen than fetal hemoglobin (HbF).
So, for example, this is important during gestation. Since a baby needs to compete
for oxygen with mom, it’s helpful to have a higher affinity for O2 than mom to help
efficiently transfer oxygen to the baby.
So in an adult with hemoglobin A, when PCO2, temperature, or 2,3-DPG are high;
or pH is low, hemoglobin affinity for oxygen decreases and more oxygen is
unloaded in the tissues.
The curve shifts to the right because now a higher PO2, or P50, is needed to
overcome these factors and saturate hemoglobin.
On the flip side, when PCO2, temperature, or 2,3-DPG are low; or pH is high; or
we’re looking at HbF instead of Hemoglobin A, hemoglobin affinity increases and
less oxygen is unloaded in the tissues.
The curve shifts to the left because a lower PO2 is needed to saturate hemoglobin.
So far, to be clear, these factors affect hemoglobin’s affinity for oxygen, but not
hemoglobin’s capacity to bind oxygen.
Meaning, hemoglobin is still only able to bind four molecules of oxygen, but how
easily it does so when there’s a certain partial pressure of oxygen depends on the
circumstances.
Now, one additional situation to consider is when there’s something like carbon
monoxide (CO) floating around which actually competes with oxygen for binding
spots on hemoglobin.
Hemoglobin’s affinity for CO is 250 times greater than that of O2.
So hemoglobin will bind CO, forming carboxyhemoglobin, more readily than it will
bind to O2.
And once CO binds, it doesn’t unload easily, making those binding spots
unavailable to oxygen.
This has two effects. First, the affinity for hemoglobin to bind O2 increases at the
sites not filled by CO.
This is an attempt for O2 to compete with CO to bind to hemoglobin but results in
less unloading of O2 in the tissues because of increased affinity.
Second, the entire shape of the curve changes from sigmoidal to one that flattens
out to a zero slope. Where the curve flattens depends on how saturated
hemoglobin is with CO.
For example, if 50% of the binding sites on hemoglobin are filled with CO, the curve
will flatten out at about 50% hemoglobin saturation with oxygen.
And as the PCO increases, and more CO binds, the curve will flatten out at lower
oxygen saturation values.

Summary
So, to recap: The oxygen-hemoglobin dissociation curve graphs the relationship
between hemoglobin saturation with oxygen and the partial pressure of oxygen in
the blood.
The sigmoidal curve to the graph is the result of positive cooperativity which
increases hemoglobin’s affinity for oxygen as more oxygen molecules bind.
But factors like PCO2, pH, temperature, 2-3-DPG, hemoglobin type, and carbon
monoxide can all affect the affinity, causing a shift in the oxygen-hemoglobin
dissociation curve and make hemoglobin more of less likely to unload oxygen in
the tissues.