Solutions to Selected End of Chapter 5 Problems

1. Protein A has a binding site for ligand X with a K d1 of 10-6 M. Protein B has a binding site for
ligand X with a Kd of 10-9 M. Which protein has a hlgher affinity for ligand X? Explain your
reasoning. Convert the Kd to Ka for both proteins.
Affinity of binding and dissociation. Protein B (K d = 10-9M) has the higher affinity because it will bind
ligand X well at a much lower concentration of X. It takes a thousand times higher amount of X to get
similar binding Protein A (Kd = 10-6 M)
Ka = 1/Kd so for Protein A with a Kd of 10-6 M, it has a Ka of 106 M-1.

3. What is the effect of the following changes on the O 2 affinity of hemoglobin? (a) A drop in the pH of
blood plasmas from 7.4 to 7.2. (b) A decrease in the partial pressure of CO 2 in the lungs from 6kPa
(holding ones breath to 2 kPa (normal). (c) An increase in the BPG level from 5mM (normal altitudes)
to 8mM (high altitudes). (d) An increase in CO from 1.0 parts per million (ppm) in a normal indoor
atmosphere to 30 ppm in a home that has a malfunctioning or leaking furnace.

Hb binding oxygen. The affinity of Hb binding O 2 is affected by pH, CO2 and 2,3 bisphosphoglycerate
(BPG): increasing any of these decreases Hb affinity for O 2.
a. Drop in pH from 7.4 to 7.2: decreased affinity (shifts the O 2 binding curve to the right).
b. Decrease in CO2 increases affinity.
c. Increase in BPG decreases affinity.
d. Increase in CO decreases affinity.

5. Membrane proteins binding a hormone: Three different proteins binding the same hormone: below
the theta ( ) is shown at different concentrations of the hormone.
Hormone Conc. nM
0.2
0.5
1
4
10
20
50

Protein 1
0.048
0.11
0.2
0.5
0.71
0.83
0.93

Protein 2
0.29
0.50
0.67
0.89
0.95
0.97
0.99

a. What is the Kd for Protein 2: By simple inspection you can see that it is 0.5 nM
(concentration of ligand that results in half of the binding sites being bound by ligand
(hormone). The Kd of the others can be gotten just as easy.
b. Which of these proteins binds hormone most tightly (highest affinity). Protein 2 is the
winner! Protein1 Kd = 4 nM, and Protein 3 Kd is 1 nM.

Prot

6. Hb cooperativity is a property of the interaction of the and subunits. It is totally lost when the
subunits are dissociated. We shall see this in Question 7 on the next page.

7. Maternal (22) and Fetal (22) Hb binding curves:

a) Which hemoglobin has a higher affinity for oxygen under physiological conditions, HbA or
HbF? Explain.
b) What is the physiological significance of the different O 2 affinities?
c) When all the BPG is carefully removed from samples of HbA and HbF, the measured O 2
saturation curves return to normal, as shown in the graph. What is the effect of BPG on the
Q affinities of fetal and maternal hemoglobin?

a. You can see that HbF binds oxygen with a higher affinity than HbA. The fetal curve is the
left, it takes less O2 to bind HbF than it does for HbA.
b. What is important is the flow of O2 in the placenta from HbA to HbF, because HbF has a
higher affinity for O2, it lowers [O2] which causes HbA, then to release its O 2.
c. Removing BPG shifts both Hbs to the left, making them almost the same, but HbA came
further. This suggests that HbA has a tighter affinity for BPG than HbF.