MBpS e MBq

2014-2015

Bioenergtics: Application Problems - Part II

1.

G = 2.3RT(pH P pH N ) + F( N P )
G=2.3 x 8.314 x 298 x (-1.3) + 96500 x (-0.110) = - 18023 Jmol-1
To pump protons against this energy, the redox reaction has to release at least the
same amount: G= -18 kJmol-1 (or more negative)
Complex I:
H+ + NADH + Q NAD+ + QH2
Eo= + 0.045 (-0.320) = 0.365 V
Go= -2 x 96.5 x 0.365 = -70.4 kJmol-1
Enough energy. N protons = 70.4/18 = 3.9 (real value 4 protons)
Complex II:
succinato + Q fumarato + QH2
Eo= + 0.045 (+0.031) = 0.014 V
Go= -2 x 96.5 x 0.014 = -2.7 kJmol-1
The energy is not enough. (real value zero protons)
Complex IV:
O2 + 2 cit c

red

+ 2 H+ H2O + 2 cit c

ox

Eo= + 0.816 (+0.235) = 0.581 V

Go= -2 x 96.5 x 0.581 = -112.1 kJmol-1
Enough energy. N protons = 112.1/18 = 6.2 (real value 2 protons).
2.

Since O2 is the final electron acceptor:

H+ + NADH + O2 NAD+ + H2O
Eo= + 0.816 (-0.320) = 1.136 V
Go= -2 x 96.5 x 1.136 = -219.2 kJmol-1
2.5 ATP / NADH
Yield =energy used to form ATP/total energy available
= 2.5 x 30.5 / 219.2 = 0.35 (yield 35%)
However, this value is not realistic because the effective G in the mitochondria is more
negative than the standard value. If we assume that the energy of the gradient is the
value obtained in the problem above, 18 KJmol-1, and that 10 H+ are transported for
each 2e- that go from NADH to oxygen =10 x 18 / 219 = 0.85. The yield for energy
conservation in the respiratory chain is much higher (85%).
3. For the sugar only the concentration gradient matters. The energy involved
concentrating the sugar 60X is G= 8.314 x 298 x ln(60) = 10144 Jmol-1
1

Teresa Catarino and Ricardo Louro

MBpS e MBq

2014-2015

It is necessary that the energy of the sodium electrochemical

[ X ]B
gradient matches this value and it must be negative to push the G = RT ln X + zF ( B A )
[ ]A
sugar against its own gradient.
For [Na]in=14 mM and [Na]out=142 mM
-10144= -5740 + F (in-out)
in-out= (-10144+5740)/96500 = -0.046 V
Under these conditions the membrane potential has to be more negative than -46 mV.
4.

a) The ratio P/O decreases

b) The energy is dissipate as heat.
c) If the concentration of uncoupler is too high the ATP may be too small to
sustain life.

5. The P/O ratio relates directly the number of ATP that are produced per
cofactor that is reoxidised in the respiratory chain. To answer this question it is
necessary to consider: i) the number of protons pumped per pair of electrons
going from NADH to O2 (10H+/2e-); ii) How many protons are involved in the
production of 3 ATP (360 rotation), which depends on the number of subunits
of the c ring (8 for mitochondria and 14 for the chloroplast).
For mitochondria P/O = 3/(8/10) = 3.75
For chloroplast P/O = 3/(14/10) = 2.14
6.

The electron donor is H2 and the final electron acceptor is sulfate:

H2 + SO42- SO32- + H2O
Eo= + 0.480 (-0.413) = 0.893 V
Energy available: Go= -2 x 96.5 x 0.893 = -172.3 kJmol-1
Energy used: Go= -172.3 x 0.56 = -96.5 kJmol-1
Considering that 30.5 kJmol-1 are necessary to phosphorylate ADP to ATP:
N moles of ATP = 96.5 / 30.5 = 3.16
In the activation of sulfate 1 mole of ATP is hydrolyzed to AMP + PPi, and PPi is
immediately hydrolyzed. This corresponds to the consumption of 2 ATP in sulfate
activation. The net balance is 3.16 - 2= 1 ATP.

7. The various complexes and transporters have to be organized according to their

reduction potentials, such that the respiratory chain is thermodynamically favourable:
-230 -180
-150

R complex C Y(solvel)

+60

+145

+300 +480
complex A Q(membrane) complex B P

To know which complexes may be involved in proton pumping it is necessary to calculate

the energy available in each of them.
Complex C: electron donor R, electron acceptor Y:
RH2 + 2Yox R + 2Yred + 2H+
Eo= - 0.150 (-0.230) = 0.080 V
Energy available: Go= -2 x 96.5 x 0.080 = -15.4 kJmol-1
A pmf = 200 mV corresponds to 19.3 kJmol-1 (96.5 x 0.200) this means that complex C
cannot pump protons.
2

Teresa Catarino and Ricardo Louro

MBpS e MBq

2014-2015

Complex A: electron donor Y, electron acceptor Quinone:

Q + 2Yred + 2H+ QH2 + 2Yox
Eo= + 0.145 (-0.150) = 0.295 V
Energy available: Go= -2 x 96.5 x 0.295 = -56.9 kJmol-1
Since the energy of the gradient is 19.3 kJmol-1, complex A has enough energy to
pump 2 protons (and there is still a lot of energy to guarantee that the whole process
is favourable).
Complex B: electron donor QH2, electron acceptor P:
QH2 + P PH2 + Q
Eo= + 0.480 (+0.145) = 0.335 V
Energy available: Go= -2 x 96.5 x 0.335 = -64.6 kJmol-1
Since the energy of the gradient is 19.3 kJmol-1, complex B has enough energy to
pump 3 protons.
Other considerations:
A, B and C are transmembrane complexes. Y, R and P are in the aqueous phase and
the quinone in the membrane. The substrates may be involved in substrate turnover
because the reactions involve protons and the quinone may be involved in a classical
redox loop. The transmembrane complexes might be conformational pumps (with any
e-/H+ stoichiometry).
One possible solution for the exercise is the one presented in the figure. But there are
other possible solutions as long as the final stoichiometry is 6H+/2e-.

The total energy available in this respiratory chain is

RH2 + P PH2 + R
Eo= +0.480 (-0.230) = 0.710 V
Go= -2 x 96.5 x 0.710 = -137 kJmol-1
Which is enough to pump 137/19.3 = 7 protons against the gradient. Since it is
translocating only 6, the chain is energetically possible.
3

Teresa Catarino and Ricardo Louro