10.

1 Cellular Respiration: pyruvate and other oxidizable substrates, they can then trans-
fer these electrons to oxygen. Aerobic respiration therefore
Maximizing ATP Yields involves oxidative pathways in which electrons are removed
from organic substrates and transferred to coenzyme carriers,
Cellular respiration, or respiration for short, dramatically
which then transfer these electrons to oxygen, accompanied
improves the metabolic energy yield per molecule of glucose.
by the generation of ATP.
With an external electron acceptor available, complete oxida-
tion of substrates to CO2 becomes possible, and ATP yields are
much higher. By “external” we mean an electron acceptor that Respiration Includes Glycolysis, Pyruvate
is not a by-product of glucose catabolism. When an “internal” Oxidation, the Citric Acid Cycle, Electron
electron acceptor such as pyruvate or acetaldehyde is used Transport, and ATP Synthesis
(as during fermentation) to accept electrons from NADH (see We will consider aerobic respiration in five stages, as shown
Figure 9-8), the by-products are not completely oxidized to CO2. in Figure 10-1. The first three stages involve substrate oxida-
As a formal definition, cellular respiration is the flow of tion and the simultaneous reduction of coenzymes, and the
electrons, through or within a membrane, from reduced coenzymes second two involve coenzyme reoxidation and the generation
to an external electron acceptor, usually accompanied by the gen- of ATP. In cells, of course, all five stages occur continuously
eration of ATP. We have already encountered NADH as the and simultaneously.
reduced coenzyme generated by the glycolytic catabolism of Stage ●1 is the glycolytic pathway (which we encountered
sugars or related compounds. As we will see shortly, two other in Chapter 9). The steps of glycolysis are the same under
coenzymes, FAD (for flavin adenine dinucleotide) and coenzyme aerobic and anaerobic conditions and result in the oxidation
Q (or ubiquinone), also collect the electrons that are removed of glucose to pyruvate. However, in the presence of oxygen,
from oxidizable organic substrates and pass them to the termi- the fate of the pyruvate is different (see Figure 9-8). Instead
nal electron acceptor via a series of electron carriers, generat- of serving as an electron acceptor, as occurs in anaerobic fer-

Chapter 10
ing ATP in the process. mentation, pyruvate is further oxidized. In stage ● 2 , pyruvate
For many organisms, including us, the terminal electron is oxidized to generate acetyl coenzyme A (acetyl CoA), which
acceptor is oxygen, which is reduced to water and, therefore, enters the citric acid cycle (stage ●
3 ). The citric acid cycle com-
the overall oxygen-requiring process is known as aerobic pletely oxidizes acetyl CoA to CO2 and conserves most of the

|
respiration. (Note that, in medical terminology at an or- energy as high-energy reduced coenzyme molecules.

Chemotrophic Energy Metabolism: Aerobic Respiration

ganismal level, respiration refers to breathing, the uptake of Stage ●4 involves electron transport, or the transfer of elec-
oxygen.) But although many chemotrophs on Earth carry out trons from reduced coenzymes to oxygen, coupled to the active
aerobic respiration, other organisms, especially some bacteria transport (pumping) of protons across a membrane. The trans-
and archaea, can use a variety of terminal electron acceptors fer of electrons from coenzymes to oxygen is exergonic and
other than molecular oxygen and thus carry out anaerobic provides the energy that drives the pumping of protons across
respiration. Examples of alternative electron acceptors and the membrane containing the carriers. This generates an elec-
their reduced forms include elemental sulfur (S/H2S), protons trochemical proton gradient across the membrane. In stage ● 5,
(H+/H2), and ferric ions (Fe3+ /Fe2+). Respiratory processes the energy of this proton gradient is used to drive ATP synthe-
that involve electron acceptors such as these require no mo- sis in a process known as oxidative phosphorylation.
lecular oxygen and are therefore anaerobic. These anaerobic Our goal in this chapter is to understand the processes fol-
respiratory processes play important roles in nutrient cycling lowing glycolysis—the complete oxidation of pyruvate in the
and in the overall energy economy of the biosphere. citric acid cycle, electron transport from pyruvate via coen-
Here, however, we will concentrate on aerobic respira- zymes to oxygen, and the formation of a proton gradient that
tion because it is the basis of energy metabolism in the aerobic drives ATP synthesis. We will begin our discussion of aerobic
world of which we and most other familiar organisms are a energy metabolism with a close look at the mitochondrion be-
part. We will focus much of our attention on the mitochon- cause of its prominent role in eukaryotic energy metabolism.
drion (Figure 10-1) because most aerobic ATP production in
eukaryotic cells takes place within this organelle. In bacterial CONCEPT CHECK 10.1
cells, the roles of the plasma membrane and cytoplasm in en- Aerobic respiration uses an external electron acceptor rather
ergy production are analogous to the roles of the mitochon- than an internal electron acceptor, as is used in anaerobic
drial inner membrane and matrix, respectively. fermentation. Explain what this means. Why does using an
external electron acceptor provide so much more energy to
Aerobic Respiration Yields Much More Energy the cell?
than Fermentation Does
With oxygen available as the terminal electron acceptor, pyru-
vate can be oxidized completely to CO2 instead of being used to 10.2 The Mitochondrion: Where
accept electrons from NADH. Therefore, aerobic respiration has
the potential of generating up to 38 ATP molecules per glucose.
the Action Takes Place
Oxygen makes all of this possible by serving as the termi- Our discussion of aerobic respiration starts with a description
nal electron acceptor, thereby providing a means for the con- of the mitochondrion because most of aerobic energy me-
tinuous reoxidation of NADH and other reduced coenzymes. tabolism in eukaryotic cells takes place within this organelle.
After these coenzyme molecules accept electrons from Because of this, the mitochondrion is often called the “energy
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 Mitochondrion

Triglycerides

CO2 2 PYRUVATE
ATP OXIDATION Hydrolysis

Glucose Pyruvate Pyruvate Acetyl CoA Fatty acids

b oxidation
1 GLYCOLYSIS
NADH CO2
2 NADH 3
CITRIC CO2
3 NADH
CYTOSOL ACID
- CYCLE
2e
FADH2 ATP

1/2 O2
MATRIX H2O
H+ H+ H+ 4 ELECTRON
+
2 e- 2H TRANSPORT
Inner membrane +
H and
Intermembrane space PROTON
H+ H+ H+ H+
+
H+ H + H+
H+ H+ H+ H+ H+ PUMPING
Outer membrane H+ H+ H+ H H+
H+ H+ H+
H+ H+ H + H+ +
H
H+
H+ Crista

H+
ADP H+ ADP
H+ ADP
+ Pi ATP + Pi H+
H+ ATP + Pi
ATP
H+
H+ 5 ATP SYNTHESIS
H+ H+ H+
H+ H+

Figure 10-1 The Role of the Mitochondrion in Aerobic Respiration. The mitochondrion plays a central
role in aerobic respiration. Most respiratory ATP production in eukaryotic cells occurs in this organelle. Oxida-
tion of glucose and other sugars begins in the cytosol with glycolysis (stage ● 1 ), producing pyruvate. Pyruvate
is transported into the mitochondrion, where it is oxidized within the matrix to acetyl CoA (stage ● 2 ), the
primary substrate of the citric acid cycle (stage ●3 ). Acetyl CoA can also be formed by b oxidation of fatty acids.
Electron transport is coupled to proton pumping (stage ● 4 ) to produce an electrochemical proton gradient
across the inner membrane of the mitochondrion. The energy of the proton gradient drives the synthesis of ATP
from ADP and inorganic phosphate (stage ● 5 ).

powerhouse” of the eukaryotic cell. Defects in mitochondrial As early as 1850, the German biologist Rudolph Kölliker
function can lead to human diseases known as mitochon- described the presence of what he called “ordered arrays of
drial myopathies. In later chapters, we will discuss additional particles” in muscle cells. Isolated particles were found to swell
aspects of mitochondrial biology, such as protein import into in water, leading Kölliker to conclude that each particle was
mitochondria, destruction of damaged mitochondria by au- surrounded by a semipermeable membrane. These particles
tophagy, their role in programed cell death, and the mitochon- are now called mitochondria (singular: mitochondrion) and are
drial genome. Here, however, we will focus on the key role of believed to have arisen when bacterial cells were engulfed by
mitochondria in cellular energy production. larger cells but survived, taking up permanent residence in
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 the host cell cytoplasm via endosymbiosis (as discussed in in muscle cells (see Figure 4-14 and chapter-opening figure).
Chapter 4). The mitochondria in these muscle cells are organized in rows
Evidence suggesting a role for this organelle in oxidative along the fibrils responsible for contraction. A similar strate-
events began to accumulate almost a century ago. In 1913, gic localization of mitochondria occurs in flagella and cilia, as
for example, Otto Warburg showed that these particles could well as in sperm tails (see Figure 4-13).
consume oxygen. However, most of our understanding of the
role of mitochondria in energy metabolism came since the Are Mitochondria Interconnected Networks
development of differential centrifugation, pioneered by Al-
Rather than Discrete Organelles?
bert Claude (see Key Technique in Chapter 4, page 120).
Intact, functionally active mitochondria were first isolated by When seen in an electron micrograph such as that in
this technique in 1948 and were subsequently shown by Eu- Figure 10-3a, mitochondria typically appear in outline as oval
gene Kennedy, Albert Lehninger, and others to be capable of structures measuring one or more micrometers in length and
carrying out not only all the reactions of the citric acid cycle, 0.5–1.0 mm across. This appearance has fostered the widely
but also electron transport and oxidative phosphorylation. accepted view that mitochondria are discrete oval-shaped en-
tities, as depicted in Figure 10-3b. However, depending on the
cell type, mitochondria can be seen in various shapes and siz-
Mitochondria Are Often Present Where es. Recent work using electron microscopy (EM) tomography,
the ATP Needs Are Greatest which allows reconstruction of a three-dimensional model
Mitochondria are found in virtually all aerobic cells of eukary- of a cellular structure from a parallel series of cross sections,
otes and are prominent features of many cell types when ex- shows that some mitochondria are not simple ovals, but have
amined by electron microscopy (Figure 10-2). Mitochondria much more complex shapes, as shown in Figure 10-4. (The
are present in both chemotrophic and phototrophic cells and technique of EM tomography is discussed in more detail in
are therefore found not only in animals but also in plants. Their Key Technique, pages 272–273.)

Chapter 10
occurrence in phototrophic cells reminds us that even photo- Calculations based on cross-sectional views of mitochon-
synthetic organisms rely on respiration to meet energy needs. dria in electron micrographs indicate that the number of mi-
The crucial role of the mitochondrion in meeting cellular tochondria per cell is highly variable, ranging from one or a
ATP needs is often reflected in the localization of mitochon- few per cell in many protists, fungi, and algae (and in some

|
dria within the cell. Frequently, mitochondria are clustered mammalian cells as well) to a few thousand per cell in some

Chemotrophic Energy Metabolism: Aerobic Respiration

in regions of cells with the most intense metabolic activity tissues of higher plants and animals. Mammalian liver cells,
and the greatest need for ATP. An especially good example is for example, are thought to contain about 500–1000 mito-
chondria each, though only a small fraction of these are seen
in a typical thin section prepared for electron microscopy, such
Mitochondria as that shown in Figure 10-2.
The notion that the mitochondrial profiles seen in elec-
tron micrographs represent discrete organelles of known size
and abundance has been challenged by the work of Hans-
Peter Hoffmann and Charlotte Avers. After examining a com-
plete series of thin sections through an entire yeast cell, these
investigators concluded that the oval profiles observed in indi-
vidual micrographs all represent slices through a single large,
extensively branched mitochondrion. Such results suggest
that the number of mitochondria present in a cell may be con-
siderably smaller than generally believed—and that the size of
a mitochondrion may be much larger than can be calculated
from individual cross-sectional profiles seen in thin-section
electron micrographs.
Further support for the concept of mitochondria as inter-
connected networks (rather than many separate organelles)
comes from studies in which intact living cells were examined
by phase-contrast microscopy or by fluorescence microscopy
using mitochondria-specific fluorescent dyes. Such investiga-
tions revealed that living cells contain large branched mito-
chondria in a dynamic state of flux, with segments of one mi-
tochondrion frequently pinching off and fusing with another
0.5 mm mitochondrion.
Figure 10-2 Mitochondria: One or Many? Mitochondria are
Most of the discussion and illustrations in this chapter
prominent features in this cross section of a rat liver cell and were will presume the conventional view of mitochondria as dis-
traditionally thought to be discrete oval structures. However, current crete entities, but keep in mind that what we regard as indi-
research suggests that these may be separate slices through one or vidual mitochondria may well be parts of a large, dynamic
more large, multilobed mitochondria (TEM). network instead, at least in some types of cells.
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 Inner and outer Rough endoplasmic
membranes reticulum

Cristae

Outer membrane

Intermembrane
space

Inner membrane

Matrix

Cristae junction

Matrix Intracristal spaces

Cristae

(a) Electron micrograph 1 mm (b) Schematic diagram

Figure 10-3 Mitochondrial Structure. (a) A mitochondrion of a bat pancreas cell as seen by electron mi-
croscopy (TEM). The cristae are formed by infoldings of the inner membrane. (b) A mitochondrion is illustrated
schematically in this cutaway view that shows the traditional “baffle” model of cristae structure. As noted in the
text, however, this model is currently being reconsidered.

The Outer and Inner Membranes Define Two membranes. The outer membrane is not a significant
Separate Compartments and Three Regions permeability barrier for ions and small molecules because
it contains transmembrane channel proteins called porins
Figure 10-3 shows both an electron micrograph and an il-
that permit the passage of solutes with molecular weights
lustration of a mitochondrion. A distinctive feature is the
up to about 5000 (see Figure 8-9). Similar proteins are
presence of two membranes, called the outer and inner
found in the outer membrane of gram-negative bacteria.
Because porins allow the free movement of small molecules
and ions across the outer membrane, the intermembrane
space between the inner and outer membranes of the mi-
tochondrion is continuous with the cytosol with respect to
small solutes. However, enzymes targeted to the intermem-
brane space are effectively confined there because enzymes
and other soluble proteins are too large to pass through the
porin channels.
In contrast to the outer membrane, the inner
membrane of the mitochondrion presents a permeability
barrier to most solutes, thereby partitioning the mitochon-
drion into two separate compartments—the intermembrane
space and the interior of the organelle (the mitochondrial ma-
trix). The inner and outer membranes are each about 7 nm
thick and are typically separated by an intermembrane space
of about 7 nm. However, in certain spots the membranes are
in contact, and it is in these regions that proteins destined for
the mitochondrial matrix pass through the two membranes
(see Figure 19-26).
At the perimeter of the mitochondrion, the portion of
the inner membrane adjacent to the intermembrane space is
Figure 10-4 EM Tomography of an Extended
known as the inner boundary membrane. In addition, the inner
Mitochondrion. This tomogram of a mouse embryonic fibroblast membrane of most mitochondria has many distinctive infold-
shows the extended shape of the intact mitochondrion. Individual ings called cristae (singular: crista) that greatly increase its
cristae, mostly lamellar, are colored separately. Note the tubular surface area. In a typical liver mitochondrion, for example,
crista (light blue) at the bottom left. the area of the inner membrane (including the cristae) is
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 about five times greater than that of the outer membrane. space, effectively creating a third, nearly enclosed region in
Because of its large surface area, the inner membrane can ac- the mitochondrion.
commodate large numbers of the protein complexes needed The relative prominence of cristae within the mitochon-
for electron transport and ATP synthesis, thereby enhancing drion frequently reflects the relative metabolic activity of the
the mitochondrion’s capacity for ATP generation. The inner cell or tissue in which the organelle is located. Heart, kidney,
membrane is about 75% protein by weight, which is a higher and muscle cells have high respiratory activities, and thus
proportion of protein than in any other cellular membrane. their mitochondria have correspondingly large numbers of
These inner membrane proteins include the transmembrane prominent cristae. The flight muscles of birds are especially
portions of proteins involved in solute transport, electron high in respiratory activity and have mitochondria that are
transport, and ATP synthesis. The cristae also provide numer- exceptionally abundant in cristae. Plant cells, by contrast,
ous localized regions, the intracristal spaces, where protons can have lower rates of respiratory activity than do most animal
accumulate between the folded inner membranes during the cells and have correspondingly fewer cristae within their
electron transport process, which we will discuss later in the mitochondria.
chapter. The interior of the mitochondrion is filled with a semi-
It was long thought that cristae were wide, flattened fluid matrix. Within the matrix are many of the enzymes
structures with broad connections to the inner bound- involved in mitochondrial function as well as DNA molecules
ary membrane, giving rise to the “baffle” model of cristae and ribosomes. In most mammals, the mitochondrial genome
structure shown in Figure 10-3b. Thanks to EM tomogra- consists of a circular DNA molecule of about 15,000–20,000
phy, however, we know that cristae can adopt a variety of base pairs that code for ribosomal RNAs, transfer RNAs, and
shapes. It is now believed that cristae in many tissues may be about a dozen polypeptide subunits of inner-membrane pro-
tubular structures that associate in layers to form lamellar teins. Mutations in mitochondrial DNA can lead to human
(layered) cristae of irregular size and shape (Figure 10-5). disease and have been associated with aging and neurodegen-
Cristae appear to have only limited connections to the in- eration. Besides the proteins encoded by the mitochondrial

Chapter 10
ner boundary membrane through small tubular openings genome, mitochondria contain numerous nuclear-encoded
known as crista junctions. The small size of these openings proteins that are synthesized on cytoplasmic ribosomes and
(approximately 20 nm) is thought to limit diffusion of mate- then imported into mitochondria (see Chapter 19).
rials between the intracristal space and the intermembrane

|Chemotrophic Energy Metabolism: Aerobic Respiration

Mitochondrial Functions Occur in or on
Specific Membranes and Compartments
Specific functions and pathways have been localized within
the mitochondrion by disruption of the organelle and frac-
tionation of the various components. Table 10-1 lists some
of the main functions localized to each compartment of the
mitochondrion.
Most of the mitochondrial enzymes involved in pyruvate
oxidation, in the citric acid cycle, and in the catabolism of fatty

Table 10-1 Localization of Metabolic Functions

Within the Mitochondrion

Membrane or Compartment Metabolic Functions

Outer membrane Phospholipid synthesis

Fatty acid desaturation
Fatty acid elongation
Inner membrane Electron transport
Proton translocation for ATP
synthesis
Oxidative phosphorylation
Pyruvate import
Fatty acyl CoA import
Metabolite transport
0.2 mm
Matrix Pyruvate oxidation
Figure 10-5 Structure of Individual Cristae. EM tomography Citric acid cycle
was used to create this three-dimensional reconstruction of three ATP synthesis
b oxidation of fats
cristae (green, light blue, dark blue) in a mitochondrion. White ar-
DNA replication
rows point to crista junctions connecting two of the cristae with the
RNA synthesis (transcription)
inner membrane (gold), making the intracristal spaces contiguous Protein synthesis (translation)
with the intermembrane space. The outer membrane is not shown.
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