Chapter 7 Hemoglobin: A Portrait of a Protein in Action

Matching Questions
Use the following to answer questions 1-10:

Choose the correct answer from the list below. Not all of the answers will be used.
a) cooperative
b) Bohr effect
c) thalassemia
d) carbamate
e) metmyoglobin
f) superoxide
g) myoglobin
h) bicarbonate ion
i) sickle-cell anemia
j) protoporphyrin
k) fetal
l) carbonic acid

1 ____________ This is a reactive oxygen species that is damaging to biological

materials.

Ans: f
Section: 7.1

2 ____________ This is the organic portion of the heme group in hemoglobin.

Ans: j
Section: 7.1

3 ____________ This is a genetic disease due to the decreased production of

one of the subunits of hemoglobin.

Ans: c
Section: 7.4
 4 ____________ This is the chemical form in which most of the carbon dioxide
is transported in the blood.

Ans: h
Section: 7.3

5 ____________ This substance is produced when carbon dioxide reacts with

water.

Ans: l
Section: 7.3

6 ____________ This type of hemoglobin is composed of two α chains and two

γ chains.

Ans: k
Section: 7.2

7 ____________ This is the molecule whose function is to store oxygen is

muscle cells.

Ans: g
Section: Introduction

8 ____________ This oxidized hemeprotein does not reversibly bind oxygen.

Ans: e
Section: 7.1

9 ____________ This type of binding is indicated by a sigmoidal-shaped binding

curve.

Ans: a
Section: 7.2

10 ____________ This condition is a result of a single point mutation in the β

chain of hemoglobin.

Ans: i
 Section: 7.4

Fill in the Blank Questions

11 Under normal conditions, the heme iron in myoglobin and hemoglobin is in

the ____________ oxidation state.
Ans: ferrous, or Fe+2 Section: 7.1

12 The ability of myoglobin to bind oxygen depends on the presence of a bound

prosthetic group called _____________.
Ans: heme Section: 7.1

13 In hemoglobin, the iron of the heme is bonded to the four nitrogens of

porphyrin and to the proximal ______________ residue of the globin chain.
Ans: histidine Section: 7.1

14 The binding of 2-3-bisphosphogycerate to hemoglobin ____________

(increases, decreases) its affinity of oxygen binding.
Ans: decreases Section: 7.2

15 The effect of pH on oxygen-binding of hemoglobin is referred to as the

_____________.
Ans: Bohr effect Section: 7.3

16 Carbon dioxide reacts with the amino terminal groups of hemoglobin to form
carbamate groups, which carry a ______________ charge.
Ans: negative Section: 7.3

17 The T-state of hemoglobin is stabilized by a salt bridge between β1 Asp 94

and the C-terminal ___________________ of the β1 chain.
Ans: histidine Section: 7.3

18 In normal adult hemoglobin, HbA, the β6 position is a glutamate residue,

whereas in sickle-cell hemoglobin, HbS, it is a ____________ residue.
Ans: valine Section: 7.4
 19 As the pressure of carbon increases, the affinity of oxygen binding to
hemoglobin ______________.
Ans: decreases Section: 7.3

20 2,3-Bisphosphoglycerate binds only to the __________________ form of

hemoglobin.
Ans: T-, or deoxy Section: 7.2

Multiple Choice Questions

21 What factor(s) influence(s) the binding of oxygen to myoglobin?

A) The concentration of bicarbonate ion, HCO3-
B) The partial pressure of oxygen, pO2
C) The concentration of hemoglobin present
D) The concentration of 2,3-BPG
E) Responses b and d
Ans: B Section: 7.1

22 Which of the following is correct concerning the differences between

hemoglobin and myoglobin?
A) Both hemoglobin and myoglobin are tetrameric proteins.
B) Hemoglobin exhibits a hyperbolic O2 saturation curve while myoglobin
exhibits a sigmoid shaped curve.
C) Hemoglobin exhibits cooperative binding of O2 while myoglobin does
not.
D) Hemoglobin exhibits a higher degree of O2 saturation at all
physiologically relevant partial pressures of O2 than does myoglobin.
E) All of the above.
Ans: C Section: 7.1

23 Which of the following is not correct concerning myoglobin?

A) The globin chain contains an extensive α-helix structure.
B) The heme group is bound to the globin chain by two disulfide bonds to
cysteine residues
C) The iron of the heme group is in the Fe+2 oxidation state.
D) The diameter of the iron ion decreases upon binding to oxygen.
E) The function of myoglobin is oxygen storage in muscle.
 Ans: B Section: 7.1

24 The structure of normal adult hemoglobin can be described as

A) a tetramer composed of four myoglobin molecules.
B) a tetramer composed of two αβ dimmers.
C) a tetramer composed of two α2 and two β2 dimers.
D) a tetramer composed of two α2 and two γ2 dimers.
E) None of these accurately describe hemoglobin.
Ans: B Section: 7.1

25 Which of the following is correct concerning fetal hemoglobin?

A) Fetal hemoglobin is composed of two α and two γ subunits.
B) Fetal hemoglobin binds 2,3-BPG more tightly than normal adult
hemoglobin.
C) Fetal hemoglobin binds oxygen less than HbA at all pO2.
D) Fetal hemoglobin does not exist in the T-form.
E) None of the above.
Ans: A Section: 7.2

26. Hemoglobin-binding of oxygen is best described as a

A) concerted model.
B) Michaelis-Menten model.
C) sequential model.
D) combination of sequential and concerted models.
E) None of the above.
Ans: D Section: 7.2

27. 2-3 Bisphosphoglycerate

A) binds in the central cavity in the T-form of hemoglobin.
B) preferentially binds to deoxyhemoglobin and stabilizes it.
C) is present in the red blood cells.
D) All of the above.
E) None of the above.
Ans: D Section: 7.2

28. What is the Bohr effect?

A) the ability of hemoglobin to retain oxygen when in competition with
myoglobin
B) the regulation of hemoglobin-binding by hydrogen ions and carbon
 dioxide
C) the alteration of hemoglobin conformation during low oxygen stress
D) All of the above.
E) None of the above.
Ans: B Section: 7.3

29. Which of the following statements is correct for hemoglobin and oxygen
transport?
A) The oxygen binds to the proximal histidine residue of the globin chain.
B) Bonding of carbon dioxide to hemoglobin molecules increases the
binding of oxygen.
C) Hemoglobin binds more oxygen as the pH is lowered.
D) Hemoglobin binds more oxygen at higher [BPG] concentrations.
E) The binding of each O2 molecule to hemoglobin increases its affinity for
the next O2 .
Ans: E Section: 7.1

30. Which of the following describes the Bohr Effect?

A) Lowering the pH results in the release of O2 from oxyhemoglobin.
B) Increasing the pressure of CO2 results in the release of O2 from
oxyhemoglobin.
C) Increasing the pH increases the T-form of hemoglobin.
D) All of the above.
E) a and b
Ans: E Section: 7.3

31. Which of the following is correct concerning the following equilibria?

CO2 + H2O H2CO3
A) An increase in the pressure of CO2 will result in a decrease of pH.
B) This reaction is catalyzed by carbonic anhydrase.
C) The H2CO3 dissociates to H+ and bicarbonate ion, HCO3-.
D) The majority of CO2 is transported to the lungs in the form of HCO3-.
E) All of the above.
Ans: E Section: 7.3

32. Carbon dioxide forms carbamate groups in proteins by reaction with

A) aspartate residues.
B) cysteine residues.
C) N-terminal amino groups.
 D) tyrosine residues.
E) heme groups.
Ans: C Section: 7.3

33. Sickle-cell anemia is caused by

A) a decreased production of α chains of hemoglobin.
B) a substitution of a Glu residue for a Phe residue at the β6 position.
C) the loss of the heme group because the proximal His is oxidized.
D) a substitution of a Val residue for a Glu residue at the β6 position.
E) a substitution of Glu residue for His at the C-terminal of the α chain.
Ans: D Section: 7.4

34. Which of the following is correct concerning the oxygenation plot of proteins
X and Y shown below?
Oxygen Binding plot

1.0
Y (fraction saturation)

0.8

0.6 X
0.4 Y

0.2

0.0
0 20 40 60 80 100
pO2 (torr)

A) Protein Y exhibits tighter oxygen-binding than protein X.

B) Protein Y corresponds to fetal hemoglobin, and protein X corresponds
to normal adult hemoglobin.
C) Protein X corresponds to fetal hemoglobin, and protein Y corresponds
to normal adult hemoglobin.
D) Protein X corresponds to myoglobin, and protein Y corresponds to
hemoglobin.
E) None of the above.
Ans: C Section: 7.2
35. Which of the following is not correct concerning the oxygenation plot of
proteins X and Y shown below?
Oxygen binding plot

1.0
Y (fraction saturation)

0.8

0.6 X

0.4 Y

0.2

0.0
0 20 40 60 80 100
pO2 (torr)

A) Protein X exhibits tighter oxygen binding than protein Y.

B) Protein Y would function as a better transport protein than protein X.
C) Protein X exhibits cooperative binding, whereas Y does not.
D) Protein X corresponds to myoglobin, and protein Y corresponds to
hemoglobin.
E) Protein Y contains multiple binding sites.
Ans: C Section: 7.1

36. Which is not correct concerning the models that are accepted to describe
cooperative binding?
A) In the sequential model, the binding of a ligand changes the
conformation of the subunit to which it binds, which in turn induces a
change in neighboring subunits.
B) All known allosteric proteins exhibit either the concerted or sequential
model exclusive of the other.
C) Both models incorporate a low affinity T-state and a higher affinity R-
state.
D) Both models explain the sigmoid-shaped binding curve.
E) In the concerted model, all molecules exist either in the T-state or the
R-state.
Ans: B Section: 7.2
37. Consider the oxygen-binding profile at three different pH values of 7.6, 7.4,
and 7.2. Which statement is most correct?

Hb Oxygen binding

1
Y (fraction saturation)

0.8

0.6 X
Y
0.4 Z

0.2

0
0 20 40 60 80 100 120
pO2 (torr)

A) Curve X most likely corresponds to pH 7.2.

B) Curve Z most likely corresponds to pH 7.6.
C) Hb has a higher affinity for oxygen at the pH of curve Z.
D) Curve Y most likely corresponds to pH 7.4.
E) pH has no effect on the oxygenation of hemoglobin.
Ans: D Section: 7.3

38. What would be the expected result of a Lys residue being substituted with a
Ser residue in the BPG binding site of hemoglobin?
A) BPG would bind tighter because of the loss of a positive charge.
B) BPG would bind tighter because of the gain of a positive charge.
C) BPG would bind less tightly because of the loss of a positive charge
D) BPG would bind less tightly because of the gain of a positive charge.
E) This substitution would have no effect on the binding of BPG.
Ans: C Section: 7.3
Short-Answer Questions

39. Why is it advantageous for hemoglobin to have allosteric properties?

Ans Hemoglobin binds oxygen in a positive cooperative manner. This allows
: it to become saturated in the lungs, where oxygen pressure is high.
When the hemoglobin moves to tissues, the lower oxygen pressure
induces it to release oxygen and thus deliver oxygen where it is needed.
Section: 7.1

40. What is fetal hemoglobin? How does it differ from adult hemoglobin?
Ans Fetal hemoglobin contains two  and two  chains, in contrast to adult
: hemoglobin with two  and two  chains. The fetal hemoglobin  chain
is probably a result of gene duplication and divergence. The difference
in the chains results in a lower binding affinity of 2-3 BPG to fetal
hemoglobin. Thus, the fetal hemoglobin has a higher affinity for oxygen,
and the oxygen is effectively transferred from the mother’s hemoglobin
to fetal hemoglobin.
Section: 7.2

41. What is metmyoglobin?

Ans Metmyoglobin is formed when the heme iron ion, which is normally in
: the +2 oxidation state, is oxidized to the +3 oxidation state. This
oxidized form of myoglobin does not bind dioxygen and is not
functional.
Section: 7.1

42. Describe the octahedral coordination sphere of the iron ion in hemoglobin
and myoglobin.
Ans The Fe+2 ion is coordinated to the four nitrogens in the center of the
: protoporphyrin of the heme. The fifth coordination site is occupied by
the “proximal histidine” of the globin chain. The oxygen is bound to the
sixth coordination site of the iron.
Section: 7.1

43. What functional role does the “distal histidine” play in the function of
myoglobin and hemoglobin?
Ans The bonding between the iron and oxygen can be described as a
: combination of resonance structures, one with Fe2+ and dioxygen and
another with Fe3+ and superoxide. The “distal histidine” donates a
 hydrogen bond to this complex stabilizing the complex and inhibits the
oxidation of the iron to the ferric state.
Section: 7.1

44. Draw the oxygen-binding curve of myoglobin and that of hemoglobin.

Indicate the partial pressure of oxygen in the lungs and the range of pressure
in tissue.
Ans
: Oxygen binding of Hb and Mb

0.8
Y (fraction saturation)

0.6
Hb
Mb
0.4

0.2
↑
↑20 – 40 torr0↑ Lungs
0 50 100
pO2 (torr)

Figure should look like Fig. 7.7

Section: 7.2

45. Describe the structure of normal adult hemoglobin.

Ans Normal adult hemoglobin, HbA, is a tetramer. It is composed of two α
: subunits and two β subunits. Each subunit has a structure very similar
to myoglobin. It can be best described as a pair of identical αβ dimers.
Each subunit contains a heme group. So, each molecule of hemoglobin
can bind up to four molecules of oxygen.
Section: 7.1

46. Briefly describe cooperative binding.

Ans Cooperative binding occurs in multi-subunit proteins that possess
: multiple binding sites. The binding of a ligand to one site causes a
 conformational change that influences the binding of the ligand to the
next site. The binding sites are not independent, but each binding
event affects the affinity of the next binding event.
Section: 7.1

47. Describe the concerted model to explain allosteric cooperative binding.

Ans The protein exists in two conformations, a T-state (for tense) that has a
: lower affinity for the ligand and an R-state (for relaxed) that has a
higher affinity for the ligand. In the concerted model, all of the
molecules exist either in the T-state or in the R-state. At each ligand
concentration, there is an equilibrium between the two states. An
increase in the ligand concentration shifts the equilibrium from the T- to
the R-state.
Section: 7.2

48. Describe the role of 2,3-bisphosphoglycerate in the function of hemoglobin.

Ans 2,3-bisphosphoglycerate, 2,3-BPG, is a relatively small, highly anionic
: molecule found in the RBC. 2,3-BPG only binds to the center cavity of
deoxyhemoglobin (T-state). The size of the center cavity decreases
upon the change to the R-form so that it cannot bind to the R-state.
Thus, the presence of 2,3-BPG shifts the equilibrium toward the T-state.
T-state is unstable, and without BPG, the equilibrium shifts so far
toward the R-state that little oxygen would be released under
physiological conditions.
Section: 7.2

49. Describe the chemical basis of the Bohr effect.

Ans The effect observed by Christian Bohr is that hemoglobin becomes
: deoxygenated as the pH decreases. In deoxyhemoglobin, three amino
acid residues form two salt bridges that stabilize the T-state. One of
these is formed between the C-terminal His β146 and an Asp residue
(β94). As the pH increases, this stabilizing salt bridge is broken because
His becomes deprotonated and loses its positive charge. At lower pH
values, this His is positively charged. The formation of the salt bridge
shifts the equilibrium from the R-state to the T-state, thus releasing
oxygen.
Section: 7.3
50. Describe how carbon dioxide affects the oxygenation of hemoglobin.
Ans Increased levels of carbon dioxide cause hemoglobin to release oxygen.
: The more active the tissue, the more fuel is burned and the more CO2 is
produced. These active tissue cells have the greatest need for oxygen
to produce more energy. The CO2 combines with the N-terminal amino
groups to form negatively charge carbamate groups. The negatively
charge carbamate groups form salt bridges that stabilize the T-state.
Thus, the increase of carbon dioxide causes the conversion of the R-
state to the T-state, releasing the bound oxygen to the tissues
producing the most CO2.
Section: 7.3

51. Briefly describe the cause of sickle-cell anemia.

Ans Sickle-cell anemia is a genetic disorder that is the result of a single
: substitution of β6 Glu with a Val. This changes a negatively charged side
chain to a nonpolar, hydrophobic side chain. This Val binds into a
hydrophobic pocket on the β chain of an adjacent molecule whose β6
Val binds to another molecule, thus hemoglobin aggregates. These
aggregates form long fibers that strain the RBC and force into a sickled
shape. The distorted red blood cells clog capillaries and impair blood
flow, resulting in the sickle-cell crisis. The sickled cells are then
destroyed, resulting in the anemia.
Section: 7.4

52. What is thalassemia?

Ans Thalassemia is caused by the substantial decreased production of one of
: the subunits of hemoglobin. In α-thalassemia, the decreased
production of the α chain results in the formation of tetramers of only
the β chain. This β4 binds oxygen more tightly than HbA and does not
exhibit cooperative binding. In β-thalassemia, the α chains form
insoluble aggregates in the immature red blood cells.
Section: 7.4

53. What is the role of α-hemoglobin stabilizing protein?

Ans Four genes express the α chains, and only two genes express the β
: chain. Thus, there is an excess of α chains, which if allowed, would
aggregate and become insoluble. Red blood cells produce α-
hemoglobin stabilizing protein (AHSP), which binds to the α chain
monomers to from a soluble complex. This prevents the aggregation of
 the α subunits.
Section: 7.4