Chapter two: part 4: cellular respiration

1. Energy in living systems

 Living organisms require free energy to power life processes such as growth,
reproduction, movement, and active transport.
 ATP (adenosine triphosphate) functions as the energy currency for cells. It
allows the cells to store energy and transfer it within the cells to provide
energy for cellular processes such as growth, movement and active transport.
 The ATP molecule consists of a ribose sugar and an adenine base with three
phosphates attached.
 In the hydrolysis of ATP, free energy is supplied when a phosphate group or
two are detached, and either ADP (adenosine diphosphate) or AMP
(adenosine monophosphate) is produced.
 Energy derived from the metabolism of glucose is used to convert ADP to
ATP during cellular respiration.
 Energy production within a cell involves many coordinated chemical
pathways. Most of these pathways are combinations of oxidation and
reduction reactions. Oxidation and reduction occur in tandem.
 An oxidation reaction strips an electron from an atom in a compound, and
the addition of this electron to another compound is a reduction reaction.
Because oxidation and reduction usually occur together, these pairs of
reactions are called oxidation-reduction reactions, or redox reactions.

Electrons and Energy

 The removal of an electron from a molecule, oxidizing it, results in a
decrease in potential energy in the oxidized compound. However, the
electron (sometimes as part of a hydrogen atom) does not remain unbonded
in the cytoplasm of a cell. Rather, the electron is shifted to a second
compound, reducing the second compound.
 The shift of an electron from one compound to another removes some
potential energy from the first compound (the oxidized compound) and
increases the potential energy of the second compound (the reduced
compound).
  The transfer of electrons between molecules is important because most of
the energy stored in atoms and used to fuel cell functions is in the form of
high-energy electrons.
 The transfer of energy in the form of electrons allows the cell to transfer and
use energy in an incremental fashion—in small packages rather than in a
single, destructive burst.
Electron Carriers
 In living systems, a small class of compounds functions as electron shuttles:
They bind and carry high-energy electrons between compounds in pathways.
 The principal electron carriers are derived from the B vitamin group and are
derivatives of nucleotides.
 These compounds can be easily reduced (that is, they accept electrons) or
oxidized (they lose electrons).
 Nicotinamide adenine dinucleotide (NAD) is derived from vitamin B3,
niacin. NAD+ is the oxidized form of the molecule; NADH is the reduced
form of the molecule after it has accepted two electrons and a proton (which
together are the equivalent of a hydrogen atom with an extra electron).

 NAD+ can accept electrons from an organic molecule according to the

general equation:
RH Reducing agent + NAD+ Oxidizing agent→ NADH Reduced +R
Oxidized
 When electrons are added to a compound, they are reduced. A compound
that reduces another is called a reducing agent. In the above equation, RH is
a reducing agent, and NAD+ is reduced to NADH.
 When electrons are removed from the compound, it is oxidized. A
compound that oxidizes another is called an oxidizing agent. In the above
equation, NAD+ is an oxidizing agent, and RH is oxidized to R.
 Similarly, flavin adenine dinucleotide (FAD+) is derived from vitamin B2,
also called riboflavin. Its reduced form is FADH2.
 A second variation of NAD, NADP, contains an extra phosphate group.
Both NAD+ and FAD+ are extensively used in energy extraction from sugars,
and NADP plays an important role in anabolic reactions and photosynthesis.
ATP in Living Systems
A living cell cannot store significant amounts of free energy. Excess free energy
would result in an increase of heat in the cell, which would result in excessive
thermal motion that could damage and then destroy the cell. Rather, a cell must be
able to handle that energy in a way that enables the cell to store energy safely and
release it for use only as needed. Living cells accomplish this by using the
compound adenosine triphosphate (ATP). ATP is often called the “energy
currency” of the cell, and, like currency, this versatile compound can be used to fill
any energy need of the cell. How? It functions similarly to a rechargeable battery.
When ATP is broken down, usually by the removal of its terminal phosphate
group, energy is released. The energy is used to do work by the cell, usually by the
released phosphate binding to another molecule, activating it. For example, in the
mechanical work of muscle contraction, ATP supplies the energy to move the
contractile muscle proteins. Recall the active transport work of the sodium-
potassium pump in cell membranes. ATP alters the structure of the integral protein
that functions as the pump, changing its affinity for sodium and potassium. In this
way, the cell performs work, pumping ions against their electrochemical gradients.
ATP Structure and Function
At the heart of ATP is a molecule of adenosine monophosphate (AMP), which is
composed of an adenine molecule bonded to a ribose molecule and to a single
phosphate group. The addition of a second phosphate group to this core molecule
results in the formation of adenosine diphosphate (ADP); the addition of a third
phosphate group forms adenosine triphosphate (ATP).

The addition of a phosphate group to a molecule requires energy. Phosphate groups

are negatively charged and thus repel one another when they are arranged in series,
as they are in ADP and ATP. This repulsion makes the ADP and ATP molecules
inherently unstable. The release of one or two phosphate groups from ATP, a
process called dephosphorylation, releases energy.

Energy from ATP

 Hydrolysis is the process of breaking complex macromolecules apart.
During hydrolysis, water is split, or lysed, and the resulting hydrogen atom
(H+) and a hydroxyl group (OH-) are added to the larger molecule.
 The hydrolysis of ATP produces ADP, together with an inorganic phosphate
ion (Pi), and the release of free energy.
 To carry out life processes, ATP is continuously broken down into ADP, and
like a rechargeable battery, ADP is continuously regenerated into ATP by
the reattachment of a third phosphate group.
  Water, which was broken down into its hydrogen atom and hydroxyl group
during ATP hydrolysis, is regenerated when a third phosphate is added to the
ADP molecule, reforming ATP.
 Obviously, energy must be infused into the system to regenerate ATP. In
nearly every living thing on earth, the energy comes from the metabolism of
glucose. In this way, ATP is a direct link between the limited set of
exergonic pathways of glucose catabolism and the multitude of endergonic
pathways that power living cells.

Phosphorylation
 Recall that, in some chemical reactions, enzymes may bind to several
substrates that react with each other on the enzyme, forming an intermediate
complex. An intermediate complex is a temporary structure, and it allows
one of the substrates (such as ATP) and reactants to react with each other
more readily; in reactions involving ATP, ATP is one of the substrates and
ADP is a product.
 During an endergonic chemical reaction, ATP forms an intermediate
complex with the substrate and enzyme in the reaction. This intermediate
complex allows the ATP to transfer its third phosphate group, with its
energy, to the substrate, a process called phosphorylation.
 Phosphorylation refers to the addition of phosphate (~P). This is illustrated
by the following generic reaction:
A + enzyme + ATP→ [A − enzyme − ∼P] → B + enzyme + ADP + phosphate ion

 When the intermediate complex breaks apart, the energy is used to modify
the substrate and convert it into a product of the reaction.
 The ADP molecule and a free phosphate ion are released into the medium
and are available for recycling through cell metabolism.
Substrate Phosphorylation
ATP is generated through two mechanisms during the breakdown of glucose. A
few ATP molecules are generated (that is, regenerated from ADP) as a direct result
of the chemical reactions that occur in the catabolic pathways.
A phosphate group is removed from an intermediate reactant in the pathway, and
the free energy of the reaction is used to add the third phosphate to an available
ADP molecule, producing ATP. This very direct method of phosphorylation is
called substrate-level phosphorylation.

Oxidative Phosphorylation
 Most of the ATP generated during glucose catabolism, however, is derived
from a much more complex process, chemiosmosis, which takes place in
mitochondria within a eukaryotic cell or the plasma membrane of a
prokaryotic cell.
 Chemiosmosis, a process of ATP production in cellular metabolism, is used
to generate 90 percent of the ATP made during glucose catabolism and is
also the method used in the light reactions of photosynthesis to harness the
energy of sunlight.
 The production of ATP using the process of chemiosmosis is
called oxidative phosphorylation because of the involvement of oxygen in
the process.
2. Glycolysis

 All organisms, from simple bacteria and yeast to complex plants and
animals, carry out some form of cellular respiration to capture and supply
free energy for cellular processes.
 Although cellular respiration and photosynthesis evolved as independent
processes, today they are interdependent. The products of photosynthesis,
carbohydrates, and oxygen gas, are used during cellular respiration.
Likewise, the byproduct of cellular respiration, CO2 gas, is used during
photosynthesis.
 Glycolysis is the first pathway used in the breakdown of glucose to extract
free energy. Used by nearly all organisms on earth today, glycolysis likely
evolved as one of the first metabolic pathways. It is important to note that
glycolysis occurs in the cytoplasm of both prokaryotic and eukaryotic cells.

 Like all metabolic pathways, glycolysis occurs in steps or stages.

o In the first stage, the six-carbon ring of glucose is prepared for

cleavage (“splitting”) into two three-carbon molecules by investing
two molecules of ATP to energize the separation. As glucose is
metabolized further, bonds are rearranged through a series of enzyme-
catalyzed steps, and free energy is released to form ATP from ADP
 and free phosphate molecules. The availability of enzymes can affect
the rate of glucose metabolism. Two molecules of pyruvate are
ultimately produced. High-energy electrons and hydrogen atoms pass
to NAD+, reducing it to NADH. Although two molecules of ATP were
invested to destabilize glucose at the beginning of the process, four
molecules of ATP are formed by substrate-level phosphorylation,
resulting in a net gain of two ATP and two NADH molecules for the
cell.

 Glycolysis is the first step in the breakdown of glucose to extract energy for
cellular metabolism. Nearly all living organisms carry out glycolysis as part
of their metabolism.
 The process does not use oxygen and is therefore anaerobic. Glycolysis
takes place in the cytoplasm of both prokaryotic and eukaryotic cells.
Glucose enters heterotrophic cells in two ways.
o One method is through secondary active transport in which the
transport takes place against the glucose concentration gradient.
o The other mechanism uses a group of integral proteins called GLUT
proteins, also known as glucose transporter proteins. These
transporters assist in the facilitated diffusion of glucose.

 Glycolysis begins with the six-carbon ring-shaped structure of a single

glucose molecule and ends with two molecules of a three-carbon
sugar called pyruvate.
 Glycolysis consists of two distinct phases.
o The first part of the glycolysis pathway traps the glucose
molecule in the cell and uses energy to modify it so that the six-
carbon sugar molecule can be split evenly into the two three-
carbon molecules.
o The second part of glycolysis extracts energy from the
molecules and stores it in the form of ATP and NADH, the
reduced form of NAD+.
First Half of Glycolysis (Energy-Requiring Steps)
Step 1. The first step in glycolysis is catalyzed by hexokinase, an enzyme with
broad specificity that catalyzes the phosphorylation of six-carbon sugars.
 Hexokinase phosphorylates glucose using ATP as the source of the
phosphate, producing glucose-6-phosphate, a more reactive form of
glucose.
 This reaction prevents the phosphorylated glucose molecule from continuing
to interact with the GLUT proteins, and it can no longer leave the cell
because the negatively charged phosphate will not allow it to cross the
hydrophobic interior of the plasma membrane.
Step 2. In the second step of glycolysis, an isomerase converts glucose-6-
phosphate into one of its isomers, fructose-6-phosphate.
 An isomerase is an enzyme that catalyzes the conversion of a molecule into
one of its isomers.
Step 3. The third step is the phosphorylation of fructose-6-phosphate, catalyzed
by the enzyme phosphofructokinase.
 A second ATP molecule donates a high-energy phosphate to fructose-6-
phosphate, producing fructose-1,6-bisphosphate.
 In this pathway, phosphofructokinase is a rate-limiting enzyme. It is active
when the concentration of ADP is high; it is less active when ADP levels are
low and the concentration of ATP is high. Thus, if there is “sufficient” ATP
in the system, the pathway slows down. This is a type of end product
inhibition, since ATP is the end product of glucose catabolism.
Step 4. The newly added high-energy phosphates further destabilize fructose-1,6-
bisphosphate. The fourth step in glycolysis employs an enzyme, aldolase, to cleave
fructose-1,6-bisphosphate into two three-carbon isomers: dihydroxyacetone-
phosphate and glyceraldehyde-3-phosphate.
Step 5. In the fifth step, an isomerase transforms the dihydroxyacetone-
phosphate into its isomer, glyceraldehyde-3-phosphate.
 Thus, the pathway will continue with two molecules of glyceraldehyde-3-
phosphate. At this point in the pathway, there is a net investment of energy
from two ATP molecules in the breakdown of one glucose molecule.
Second Half of Glycolysis (Energy-Releasing Steps)
So far, glycolysis has cost the cell two ATP molecules and produced two small,
three-carbon sugar molecules. Both of these molecules will proceed through the
second half of the pathway, and sufficient energy will be extracted to pay back the
two ATP molecules used as an initial investment and produce a profit for the cell
of two additional ATP molecules and two even higher-energy NADH molecules.
Step 6. The sixth step in glycolysis oxidizes the sugar (glyceraldehyde-3-
phosphate), extracting high-energy electrons, which are picked up by the electron
carrier NAD+, producing NADH. The sugar is then phosphorylated by the addition
of a second phosphate group, producing 1,3-bisphosphoglycerate. Note that the
second phosphate group does not require another ATP molecule.
 The continuation of the reaction depends upon the availability of the
oxidized form of the electron carrier, NAD+. Thus, NADH must be
continuously oxidized back into NAD+ in order to keep this step going.
 If NAD+ is not available, the second half of glycolysis slows down or stops.
If oxygen is available in the system, the NADH will be oxidized readily,
though indirectly, and the high-energy electrons from the hydrogen released
in this process will be used to produce ATP.
 In an environment without oxygen, an alternate pathway (fermentation) can
provide the oxidation of NADH to NAD+.
Step 7. In the seventh step, catalyzed by phosphoglycerate kinase (an enzyme
named for the reverse reaction), 1,3-bisphosphoglycerate donates a high-energy
phosphate to ADP, forming one molecule of ATP. (This is an example of
substrate-level phosphorylation.)
A carbonyl group on the 1,3-bisphosphoglycerate is oxidized to a carboxyl group,
and 3-phosphoglycerate is formed.
Step 8. In the eighth step, the remaining phosphate group in 3-phosphoglycerate
moves from the third carbon to the second carbon, producing 2-phosphoglycerate
(an isomer of 3-phosphoglycerate). The enzyme catalyzing this step is a mutase
(isomerase).
Step 9. Enolase catalyzes the ninth step. This enzyme causes 2-phosphoglycerate
to lose water from its structure; this is a dehydration reaction, resulting in the
formation of a double bond that increases the potential energy in the remaining
phosphate bond and produces phosphoenolpyruvate (PEP).
Step 10. The last step in glycolysis is catalyzed by the enzyme pyruvate kinase
(the enzyme in this case is named for the reverse reaction of pyruvate’s conversion
into PEP) and results in the production of a second ATP molecule by substrate-
level phosphorylation and the compound pyruvic acid (or its salt form, pyruvate).
 Many enzymes in enzymatic pathways are named for the reverse reactions,
since the enzyme can catalyze both forward and reverse reactions.
Outcomes of Glycolysis
 Glycolysis starts with glucose and ends with two pyruvate molecules, a total
of four ATP molecules and two molecules of NADH.
 Two ATP molecules were used in the first half of the pathway to prepare the
six-carbon ring for cleavage, so the cell has a net gain of two ATP molecules
and 2 NADH molecules for its use. If the cell cannot catabolize the pyruvate
molecules further, it will harvest only two ATP molecules from one
molecule of glucose.
 Mature mammalian red blood cells are not capable of aerobic respiration—
the process in which organisms convert energy in the presence of oxygen—
and glycolysis is their sole source of ATP. If glycolysis is interrupted, these
cells lose their ability to maintain their sodium-potassium pumps, and
eventually, they die.
 The last step in glycolysis will not occur if pyruvate kinase, the enzyme that
catalyzes the formation of pyruvate, is not available in sufficient quantities.
In this situation, the entire glycolysis pathway will proceed, but only two
ATP molecules will be made in the second half. Thus, pyruvate kinase is a
rate-limiting enzyme for glycolysis.