O2 Transport

Linda Costanzo, Ph.D.

OBJECTIVES:

After studying this lecture, the student should understand:

1. How oxygen is carried in blood.

2. Features of adult hemoglobin and its variants.
3. Features of the oxygen-hemoglobin dissociation curve, including the sigmoidal
shape and P50.
4. The meaning of right- and left-shifts of the oxygen-hemoglobin dissociation curve
and what causes each.
5. The effect of carbon monoxide on the oxygen-hemoglobin dissociation curve.
6. How to calculate the oxygen content of blood.

I. FORMS OF O2 IN BLOOD

In the blood, O2 is carried in two forms: dissolved O2 and O2 bound to

hemoglobin (called O2- hemoglobin).

Dissolved O2. Dissolved O2 constitutes 2% of the total O2 content of the blood.

The quantity of O2 dissolved in blood is described by Henry’s law, which states
that the concentration of dissolved gas is proportional to its partial pressure; the
proportionality constant is the solubility. The solubility of O2 is 0.003 ml O2/100
ml blood/mm Hg. Thus, for an arterial PO2 of 100 mm Hg, the dissolved O2
content is 0.3 ml O2/100 ml blood. Alone, dissolved O2 is woefully inadequate for
O2 delivery to the tissues; we must have O2-hemoglobin!

O2-hemoglobin. The remaining 98% of O2 in blood is O2-hemoglobin, the major

topic of this lecture.

II. HEMOGLOBIN

A. Features of hemoglobin molecule

Hemoglobin, a globular protein, has four subunits. Each subunit has a

heme moiety (an iron-binding porphyrin) and a polypeptide chain. Adult
hemoglobin (called HbA) has two α chains and two β chains, thus it is
designated α2 β2. Each subunit can bind one molecule of O2; thus a
hemoglobin molecule can bind a total of four O2, which is referred to as
100% saturation. In the oxygenated form, hemoglobin is called
 oxyhemoglobin; in the unoxygenated form, it is deoxyhemoglobin. To
bind O2, iron must be in the ferrous state, Fe2+.

B. Variants of hemoglobin

1. Adult hemoglobin (HbA). See above.

2. Fetal hemoglobin (HbF). In fetal hemoglobin, HbF, the two β
chains are replaced by γ chains, thus it is described as α2 γ2. This
modification to the hemoglobin molecule results in a higher
affinity for O2 (than HbA), which lowers the PO2 (free, dissolved
O2) and facilitates movement of O2 from the maternal to the fetal
circulation. Within the first year of life HbF is replaced by HbA.
3. Methemoglobin. In methemoglobin, iron is in the ferric (Fe+3)
state instead of the normal Fe2+ state. Methemoglobin does not
bind O2. Methemoglobinemia is caused by drugs that oxidize Fe2+
to Fe+3, (e.g., sulfonamides) or congenital enzyme deficiency of
methemoglobin reductase (the red blood cell enzyme that normally
keeps iron in its Fe2+ state).
4. Hemoglobin S (HbS). HbS is the hemoglobin variant that causes
sickle cell disease. In HbS, the α subunits are normal and the β
subunits are abnormal, so it is designated αA2 βS2. The
deoxygenated form of αA2 βS2 forms sickle-shaped rods that distort
the shape of the red blood cells (i.e., “sickling”) and causes them to
occlude small blood vessels. αA2 βS2 has a lower affinity for O2
than does normal HbA.

III. O2-HEMOGLOBIN DISSOCIATION CURVE

Figure 1.
 O2 binds reversibly to the heme groups on hemoglobin. Therefore, each
hemoglobin molecule has the capacity to bind four molecules of O2. The percent
saturation (“O2 sats”) tells what percent of heme groups are bound to O2. When
all four hemes are bound to O2, there is 100% saturation, when three hemes are
bound to O2, there is 75% saturation, etc. The O2-hemoglobin dissociation curve
shows the relationship between % saturation and PO2 of the blood. This is one of
the most famous and most important curves in all of physiology! Live it, love it!

For convenience, the table below gives various values of PO2 and the
corresponding % saturation for the normal O2-hemoglobin dissociation curve.

PO2 % Saturation
10 25%
20 35%
25 50% (P50)
30 60%
40 75% (mixed venous blood)
50 85%
60 90%
80 96%
100 98% (≈ 100% arterial blood)

A. Sigmoidal shape

The O2-hemoglobin dissociation curve has a sigmoidal shape. Thus, rather

than a linear relationship between % saturation and PO2, the % saturation
increases steeply between a PO2 of zero and 40 mm Hg, then increases less
steeply between 40 and 60 mm Hg, and then is nearly flat between 60 and 100
mm Hg. (Note that one of the implications of the nearly “flat” portion of the
curve between 60 and 100 mm Hg is that changes in arterial PO2 in this range
have little effect on the % saturation of hemoglobin and therefore the amount
of O2 carried in the blood.)

The sigmoidal shape results from positive cooperativity of O2 binding to

hemoglobin. As each successive O2 binds, it increases the affinity for the next
O2. Binding of the first O2 increases the affinity for the second O2, etc.
Affinity for the fourth (last) O2 is the highest, which corresponds to the
portion of the curve where % saturation is near or at 100%.

B. P50

By definition, P50 is the PO2 that corresponds to 50% saturation. Changes in

P50 reflect changes in the affinity of hemoglobin for O2. Increases in P50
reflect decreased affinity. Decreases in P50 reflect increased affinity.
 C. Right- and left-shifts of the O2-hemoglobin dissociation curve

Figure 2.

Changes in affinity of hemoglobin for O2 produce changes in the P50 and shift
the O2-hemoglobin dissociation curve to the right or left.

Right-shifts of the O2-hemoglobin dissociation curve occur when there is a

decreased affinity of hemoglobin for O2, which produces an increase in P50.
When affinity is decreased, unloading of O2 is facilitated. The factors that
cause a right-shift make sense in terms of this decreased affinity and include:

1. Increases in PCO2 and decreases in pH, such as those occurring when

there is increased metabolic activity in a tissue (e.g., during exercise.
Called the Bohr effect.
2. Increases in temperature, such as during exercise.
3. Increases in 2,3-diphosphoglycerate (2,3-DPG). 2,3 DPG binds to the
β chains of deoxyhemoglobin and reduces their affinity for O2. During
hypoxia (such as at high altitude), 2,3 DPG production in red cells
increases, causing a helpful decrease in the affinity of hemoglobin for O2
(facilitates O2 unloading in tissues).
 Left-shifts of the O2-hemoglobin dissociation curve occur when there is
increased affinity of hemoglobin for O2, which produces a decrease in P50.
When affinity is increased, unloading of O2 is more difficult. The factors
causing a left-shift are:

1. Decreases in PCO2 and increases in pH, such as when there is decreased

metabolic activity in a tissue.
2. Decreases in temperature.
3. Decreases in 2,3-DPG.
4. Hemoglobin F (HbF). The mechanism of the left-shift with HbF relates to
2,3 DPG, which binds less avidly to the γ chains of HbF than to the
β chains of HbA. With less 2,3 DPG bound to HbF, the O2 affinity
increases, lowering the PO2 of the fetus and facilitating O2 diffusion from
mother to fetus.
5. Carbon monoxide (CO) not only causes a left-shift, but also a decrease in
O2-binding capacity (see below).

D. CO poisoning

Figure 3.
 Carbon monoxide (CO) binds to hemoglobin (to form carboxyhemoglobin)
with an incredibly high affinity, 250 times that of O2! Any heme groups bound
to CO cannot bind to O2, so CO poisoning decreases the O2-binding capacity
of hemoglobin. In the figure, for illustration, the O2-binding capacity of
hemoglobin was reduced to 50% of normal, meaning ½ of the heme sites were
occupied by CO. CO also increases the affinity of hemoglobin for whatever
O2 is bound (a left-shift of the O2-hemoglobin dissociation curve). Thus, the
effects of CO poisoning are devastating for O2 delivery to tissues: less O2 is
bound and the O2 that is bound is less readily released.

E. O2 Content of Blood

Blood flow and O2 content of blood are the major factors determining O2
delivery to tissues. O2 content of blood is comprised of dissolved O2 and
O2-hemoglobin.

1. Dissolved O2 is easy — it is described by Henry’s law as the PO2 times

the solubility of O2 in blood.

Dissolved O2= PO2 x solubility

P x 0.003 ml O2/100 ml
= O2
blood/mm Hg

Thus, at the normal arterial PO2 of 100 mm Hg:

100 mm Hg x 0.003 ml O2/100 ml

Dissolved O2 =
blood/mm Hg
= 0.3 ml O2/100 ml blood, or 0.3 vol %

2. O2 bound to hemoglobin depends upon three things:

a. Hemoglobin concentration. The normal value is 15 g/100 ml

of blood.
b. O2-binding capacity. The maximal amount of O2 that can be
bound to hemoglobin when there is 100% saturation of heme
sites. The O2-binding capacity is normally 1.34 ml O2/g
hemoglobin.
c. % Saturation. % of heme groups bound to O2. Varies from
zero to 100%

Thus, if the hemoglobin concentration of blood is 15 g/100 ml, the O2-

binding capacity of that hemoglobin is 1.34 ml O2/g, and the
 % saturation of arterial blood is 100% (at an arterial PO2 of 100 mm
Hg):

O2-hemoglobin = 15 g/100 ml x 1.34 ml O2/g x 100%

20.1 ml O2/100 ml blood, or 20.1 vol
=
%

If the % saturation of hemoglobin is less than 100% then, accordingly,

there will be less O2-hemoglobin.

3. Now, to calculate the total O2 content of blood at a PO2 of 100 mm Hg.

O2 content of blood = O2-hemoglobin + dissolved O2

20.1 ml O2/100 ml + 0.3 ml O2/100
=
ml
= 20.4 ml O2/100 ml, or 20.4 vol%

**Special note: ml gas/100 ml is also called “volume %.” Thus, an O2

content of 20.4 ml O2/100 ml blood is called “20.4 volume %” for short.
 4. Summary of O2 transport

Figure 4.

Humidified tracheal air has a PO2 of 150 mm Hg. Alveolar air has a lower
PO2 of 100 mm Hg because O2 has diffused from alveolar gas into
pulmonary capillary blood. Pulmonary capillary blood, which becomes
systemic arterial blood, equilibrates with alveolar gas, so it too has a PO2
of 100 mm Hg. The PaO2 of 100 mm Hg corresponds to 100% saturation
of hemoglobin on the O2-hemoglobin dissociation curve. The O2 content
of systemic arterial blood is the sum of dissolved O2 and O2-hemoglobin
per our discussion above. Dissolved O2 was 0.3 vol% and O2-hemoglobin
was 20.1 vol% for a grand total of 20.4 vol% in systemic arterial blood. In
the tissues, O2 diffuses from the capillaries to the tissues for aerobic
metabolism. Thus mixed venous blood has as lower PO2 of 40 mm Hg, a
correspondingly lower % saturation of 75% (read it off the O2-hemoglobin
curve!), and a correspondingly lower O2 content of 15 vol %. Thus 5 vol
% of O2 must have been transferred to the tissues. Mixed venous blood
will be re-loaded with O2 in the next pass through the lungs.
IV. PRACTICE QUESTIONS

1. A person who is hypoxemic has an arterial PO2 of 75 mm Hg and a venous

PO2 of 30 mm Hg. This person has a normal hemoglobin concentration and
a normal O2-binding capacity. Hemoglobin is 85% saturated at 75 mm Hg
and 55% saturated at 30 mm Hg. For arterial and venous blood, calculate
dissolved O2, O2-hemoglobin, and total O2 content. What was O2
consumption by the tissues (in vol %)?

2. What is the effect of increased PCO2, increased pH, and CO on O2-binding

capacity, P50, and affinity of hemoglobin for O2?

3. A right-shift of the O2-hemoglobin curve is:

A. Associated with a decrease in P50.

B. Associated with an increase in O2 content of blood at a PO2 of 50
mm Hg.
C. Caused by an increase in blood pH.
D. Caused by an increase in 2,3 DPG concentration.

4. Which of the following causes a decrease in O2-binding capacity of

hemoglobin?

A. Decreased hemoglobin concentration

B. CO poisoning
C. Decrease in arterial PO2 to 60 mm Hg
D. Increase in arterial PO2 to 120 mm Hg
E. Left-shift of the O2-hemoglobin curve

EXPLANATIONS

1.
2.

3. Answer = D. Start by drawing a normal O2-Hb curve and a right-shifted curve.

Write down what you know: associated with increased P50, decreased affinity, and
decreased O2 content at a given PO2; caused by increased PCO2, decreased pH,
and increased 2,3 DPG. I hope you see why choice B is incorrect – at a PO2 of 50
mm Hg, the right-shifted curve has a lower % saturation, thus a lower O2 content.
BTW: “associated with” means any association, cause or effect.

4. Answer = B. Get oriented! What is O2-binding capacity of hemoglobin? It is how

many ml of O2 each gram of hemoglobin can hold at 100% saturation. Decreased
hemoglobin concentration does not affect the amount of O2 each gram of
hemoglobin can hold. Decreasing or increasing PO2 changes the % saturation of
hemoglobin, but does not affect the amount of O2 that hemoglobin can hold at
100% saturation. Same with a left-shift of the curve; it increases the % saturation
at a given PO2 and increases affinity, but does not alter the maximum amount of
O2 that hemoglobin can hold. CO attaches to binding sites on hemoglobin and
prevents binding of O2, which reduces the O2-binding capacity of hemoglobin. I
suspect a few people learned something from this question!