B io Factsheet

www.curriculum-press.co.uk Number 175

Haemoglobin: Structure & Function

Haemoglobin is the oxygen – carrying pigment in red blood cells. 1. Oxygen diffuses across the alveoli into the haemoglobin in red cells
It is a globular protein consisting of four tightly packed polypeptide 2. Haemoglobin has a high affinity for oxygen in the high oxygen
chains. concentration that exist there
There are two identical alpha chains and two identical beta chains The point is that haemoglobin doesn’t have a high affinity no
(Fig 1). matter what the oxygen concentration is! If it did, it would never
let go of the oxygen!
Fig 1. Molecule of human haemoglobin. 3. Oxygen atoms combine with the haem groups of the
α-polypeptide chain Haem group β-polypeptide chain haemoglobin. The addition of one oxygen molecule to the first
haem group distorts the shape of the haemoglobin molecule,
making it easier for the second oxygen molecule to combine with
haem. This, in turn, makes it easier for the third oxygen molecule
to combine with a third haem group. It then becomes a little
harder for the fourth oxygen molecule to join the fourth haem
group. This trend explains the sigmoid shape of the oxygen
dissociation curve.
4. Oxyhaemoglobin is formed;
5. Haemoglobin unloads / has a low affinity for oxygen in areas eg
the respiring tissues, where oxygen is in low concentration;
6. This is a consequence of its dissociation curve i.e. small changes
in concentration gives large changes in saturation;
7. Respiration in tissues gives high CO 2 concentration, high
temperature and high H+ concentration
9. Under these conditions the dissociation curve shifts to the
β-polypeptide chain α-polypeptide chain right (Fig 2)
Fig 2. The effect of pH on the ODC (The Bohr Effect)
• The hydrophobic parts of the chains point inwards towards the
centre of the molecule and interactions between these 100
hydrophobic parts help to maintain the 3-D shape
• The hydrophilic parts point outwards and help maintain 1
saturation of Hb with oxygen (%)

haemoglobin’s solubility 80
74 2 1 pH 7.6
• Each polypeptide chain contains an iron-containing haem group
• Each haem group can bind with one oxygen molecule, so each 2 pH 7.4
60
molecule of haemoglobin can carry 4 oxygen molecules at a time. 59 3 3 pH 7.2
• The aggregation of several (4) polypeptide chains gives
haemoglobin a quaternary structure. 40
38

Typical Exam Questions

Q1. Use Fig 1 to explain why the haemoglobin molecule has a
20
quaternary structure.
Answer: It is made of several/ four polypeptides; Note it has
nothing to do with the fourth dimension of protein structure pO2/kilopascals
0 2 4 6 8 10 12 14
Q2. Why do all human haemoglobin molecules always have
At a ppO2 of 4 kilopascals, how saturated is the haemoglobin at:
identical tertiary structure?
pH 7.6 ?…………………answer = 74%
Answer: Because they contain an identical sequence of amino pH 7.4 ?…………………answer = 59%
acids and thus the bonds always occur in the same positions pH 7.2 ?…………………answer = 38%
You can see that the curve has moved to the right
By far the most common exam question concerns how haemoglobin As the tissues get more acidic, haemoglobin releases more oxygen
picks up oxygen in the lungs and offloads it at the respiring tissues. (becomes less saturated). Increasing temperature and increasing
To get good marks, precision is vital. Here are the key facts to learn. CO2 concentrations have just the same effect.
These are the conditions in the respiring tissues (high temperatures
and CO2 from respiration, high H+ concentrations from dissociation
of bicarbonate ions or production of lactic acid.
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Extract Chief Examiner’s Report You can see that at low partial pressures of oxygen (as there are in the
Many candidates had a poor understanding of the processes involved in placenta), the foetal haemoglobin is much more saturated than the maternal
the transport of oxygen by haemoglobin and the dissociation curve. It haemoglobin. This is because fetal haemoglobin has a greater affinity for
was often stated that haemoglobin has ‘ a high affinity for oxygen’, but oxygen at low partial pressures of oxygen than adult haemoglobin.
rarely was the answer qualified by explaining that it has a high affinity at
At the low partial pressure of oxygen at the placenta the maternal
higher partial pressures, while oxyhaemoglobin readily dissociates at
oxyhaemoglobin dissociates and releases oxygen. The fetal circulation rapidly
lower ones. Only a small minority linked their explanation to the
carries the oxygenated blood away. There is a higher partial pressure of
dissociation curve.
oxygen in maternal blood than in fetal blood entering placenta and so the
A significant minority confused haemoglobin with red cells, and referred oxygen concentration gradient is maintained. It is the ability of the foetal
to oxygen being transported in the ‘dip’ in the haemoglobin. haemoglobin to pick up oxygen at the partial pressures at which the maternal
haemoglobin is dissociating that is crucial.
The commonest problem was the use of inappropriate terminology. For
example, descriptions of the haemoglobin ‘picking up’ or ‘dropping off’
Extract from Chief Examiner’s report
oxygen were frequent. When describing the formation of oxyhaemoglobin,
Many candidates explained that fetal haemoglobin has a high affinity
candidates frequently made statements such as ‘haemoglobin joins with
for oxygen, but only the better candidates went on to explain that this
4 oxygen molecules’.
means that fetal haemoglobin takes up oxygen when adult oxygen
Fig 3 shows the oxygen dissociation curve of haemoglobin from a mammal haemoglobin dissociates.
at two different temperatures, 38 ºC and 43 ºC.
Structure of foetal and maternal haemoglobin
Fig 3. Foetal red cells are nucleated and contain foetal haemoglobin, which is a
100 38oC conjugated protein with four haem prosthetic groups linked to two α and
90
two γ polypeptide chains. Thus their different affinities for oxygen have a
80 structural basis. One difference between them is shown below.
43oC
70
Sequence of amino acids in part of adult haemoglobin
60
- Phe – Ala – Thr – Leu – Ser – Glu – Leu – His – Cys –
50
40 Sequence of amino acids in corresponding part of fetal haemoglobin
30 - Phe – Ala – Gln – Leu – Ser – Glu – Leu – His – Cys –
20 The difference in the sequence of amino acids results in different binding
10 sites for oxygen and, in turn, different oxygen dissociation curves.
0
0 2 4 6 8 10 12 14 Another difference between the blood of the mother and the foetus is that
Partial pressure of oxygen /kPa foetal red cells are nucleated. As birth approaches, foetal haemoglobin
genes are switched off, adult haemoglobin genes are
You can see that, at any particular partial pressure of oxygen, haemoglobin switched on and adult red cells containing adult haemoglobin are produced.
is less saturated at 43 ºC than at 38 ºC. that is, if the temperature increases,
the haemoglobin releases more of its oxygen. Effect of altitude
Blood that is fully (100%) saturated with oxygen carries 105 cm of oxygen 3 Less oxygen is available at high altitudes. When humans live at high altitudes,
in 1 dm3(litre) of blood. Imagine 1 dm3 of blood that has become 90% the concentration of DPG (2,3-diphosphoglycerate, in their red blood
saturated at 38 ºC. It now reaches a part of the body where intense cells increases. DPG is a substance which affects the dissociation curve.
respiration is occurring. Oxygen is being used up rapidly in respiration to Fig 5 shows the effect of a high concentration of DPG on the dissociation
release ATP. This metabolic activity has increased the temperature. The Fig 5. Effect of DPG on dissociation curve
temperature is 43ºC and the partial pressure of oxygen is 4 kPa. At this
temperature and pressure, haemoglobin will become 18% saturated (see 100 low concentration of DPG
Fig 3). The difference ( 90% -18% =72% and 72% of 105 = 75.6cm3 will
% saturation of haemogloblin

have been released to the respiring tissues.

80
Mother and foetus
with oxygen

The foetus needs to get its oxygen from the mother’s blood. Differences in 60
maternal and foetal haemoglobin ensure that this happens.
Fig 4 shows the dissociation curves for fetal and maternal haemoglobin. High concentration of DPG
40

Fig 4 100 Fetal haemoglobin 20

% saturation of haemoglobin

Maternal haemoglobin
0
0 2 4 6 8 10 12 14
with oxygen

Partial pressure of oxygen /kPa

50
curve.
At low partial pressures of oxygen, eg 2kPa, the haemoglobin with high
DPG in the red cells has released 7-8 % more oxygen than when there was
a low DPG concentration. Once again the haemoglobin has releasedoxygen
0
Partial pressure of oxygen

2
175 Haemoglobin: Structure & Function Bio Factsheet
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just when it is most needed. 4. The graph shows the dissociation curve for adult human
Practice Questions haemoglobin.
1. The graph shows the dissociation curve for human
100
oxyhaemoglobin.

saturation of haemogloblin
Using the graph, explain how haemoglobin transports oxygen 90
80

with oxygen /%
100
70
60
80
% saturation of

50
haemogloblin

60 40
30
40 20
10
20 0
0 2 4 6 8 10 12 14 16
In respiring tissue In lungs
0
4 6 8 0 10 2
12 Partial pressure of oxygen
Partial pressure of oxygen /kPa
from the lungs to respiring tissues. (5 marks) (a) Blood leaving the lungs is 98% saturated. 1 dm3 blood
leaving the lungs carries 100 cm3 oxygen. Use the graph to
2 The graph shows a dissociation curve for human haemoglobin
calculate the amount of oxygen that this volume of blood
at pH 7.4. pH 7.4
100 will unload to the respiring tissue. (2 marks)
haemogloblin with oxygen

(b) Outline the conditions in the tissues that stimulate

oxyhaemoglobin to offload oxygen (3 marks)
% saturation of

Answers
50 1. Haemoglobin has high affinity for oxygen at high partial pressures in
alveoli;
Becomes saturated;
Oxygen molecules bind to haem groups;
In tissues, oxygen partial pressure much lower;
Haemoglobin has low affinity in these conditions;
0 Oxygen offloaded;
Partial pressure of oxygen
Steep curve reflects large reduction in saturation for small drop in
partial pressure;
(a) (i) Sketch a curve on the graph to show the likely position 2. (a) (i) curve to right of curve for pH 7.4; 1
of the dissociation curve at pH 7.2. (1 mark) (ii) aerobic respiration produces carbon dioxide;
(ii) Suggest what might cause this change in pH (3 marks) carbon dioxide dissolves in blood;
(iii) Explain how this change in pH affects the supply of forming acid;
oxygen by haemoglobin to respiring tissues. (2 marks) increases hydrogen ion concentration;
(iii) more oxygen unloaded/given up / affinity decreased /reduced
3. The graph shows the dissociation curves for fetal and maternal saturation;
oxyhaemoglobin dissociates at higher oxygen concentration/
100 Fetal haemoglobin partial pressure / more oxygen unloaded at the same ppO2;
% saturation of haemoglobin

Maternal haemoglobin 3 (a) High pp of oxygen in lungs, low pp in placenta;

High percentage of haemoglobin can bind / saturate with oxygen in
lungs;
with oxygen

Low percentage of haemoglobin can bind / saturate with oxygen in

placenta;
50 Dissociates in placenta;
(b) Fetal haemoglobin has a higher affinity for oxygen / eq.;
At low pps of oxygen;
Fetal haemoglobin can associate with oxygen at low partial
pressures;
Maternal haemoglobin dissociates at low partial pressures
0
Partial pressure of oxygen 4. (a) Blood is 0.98 x = 100cm3 x = 100/0.98 = 102cm3;
Therefore 40% of 102cm3 = 40-41%;
haemoglobin (b) higher temperature;
(a) Use the graph to explain how maternal haemoglobin can greater H+ concentration;
load oxygen in the lungs and unload oxygen in the placenta.
(3 marks)
Acknowledgements: This Factsheet was researched and written by Kevin Byrne.
(b) How does fetal haemoglobin makes it possible for the fetus Curriculum Press, Bank House, 105 King Street, Wellington, Shropshire, TF1
to obtain oxygen from the mother’s blood ? (2 marks) 1NU. ISSN 1351-5136
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