Chapter 10 Chemotrophic Energy Metabolism: Aerobic Respiration

Anatomy of Mitochondria

Composed of inner and outer mitochondrial membrane

Inner membrane infolded into cristae
Between two membranes is intermembrane space
Outer mitochondrial membrane contains several enzymes, including monoamine oxidase,
which degrades neurotransmitters.
Outer membrane is relatively permeable to salts, sugars, nucleotides, coenzymes, probably
due to porins, transmembrane proteins that allow molecules < 10,000 daltons to pass -->
intermembrane space is equivalent to cytosol
Mitochondrial matrix contains all but one of the enzymes of the Krebs cycle (succinate
dehydrogenase)
Also contains DNA, RNA, ribosomes, enzymes --> involved in nucleic acid and protein
synthesis within mitochondrion.
Inner mitochondrial membrane is site for components of respiratory chain and oxidative
phosphorylation.
Membrane is also site for synthesis of steroid hormones.
First step in cleavage of cholesterol performed by enzyme localized in the inner
mitochondrial membrane.

How Mitocondria Capture Energy

Krebs cycle or citric acid cycle
Energy budget for Krebs cycle:
1 ATP produced
1 FADH2 produced
3 NADH produced (+1 from pyruvate to acetyl CoA step)
2 CO2 produced (+1 from pyruvate to acetyl CoA step)

Regulation of TCA Cycle

Several enzymes are allosterically regulated:
Pyruvate dehydrogenase complex
Isocitrate dehydrogenase
α -ketoglutarate dehydrogenase complex
Malate dehydrogenase

Pyruvate Dehydrogenase Complex

Series of three enzymes that make up complex
Allosterically inhibited by: acetyl CoA, NADH, ATP
Allosterically activated by: CoA, NAD+, AMP

Isocitrate Dehydrogenase
Allosterically inhibited by NADH
Allosterically activated by ADP

1
α -ketoglutarate Dehydrogenase
Allosterically inhibited by NADH, succinyl CoA

Malate Dehydrogenase
Allosterically inhibited by NADH

β -oxidation
Sequential breakdown of fatty acids to produce acetyl CoA
Occurs when fats are used as a source of energy
Important in migrating animals and hibernating animals

Fate of electron carriers

NADH + 1/2 O2 + H+ --> NAD+ + H20

FADH2 + 1/2 O2 ---> FAD + H20

Respiratory chain
Composed of 4 molecules:
Cytochromes
Iron-sulfur proteins
Flavoproteins
Ubiquinones

Cytochromes
Contain heme prosthetic groups and a porphyrin ring with central iron atom
Iron atom is reduced from Fe3+ to Fe2+ with acceptance of 1 electron
Several cytochromes exist and differ by size, absorption spectra, and type of heme ring
they contain
Types are: b, c, c1, a, and a3
Except for cytochrome c, all exist as integral membrane proteins
Cytochrome c is a peripheral membrane protein facing intermembrane space

Iron-sulfur Proteins
Have iron bound to sulfur atoms from cysteine

Flavoproteins
Enzymes that contain FAD or FMN as prosthetic group
Participate in re-dox reactions that involve 2 e- and 2 H+ transfers

Ubiquinones
Lipid molecules that are also electron carriers
Participate in re-dox reactions that involve 2 e- and 2 H+ transfers

Respiratory Chain

2
Respiratory chain is organized into 4 multiprotein complexes
respiratory complex I or NADH dehydrogenase - catalyzes transfer of e- from NADH
to ubiquinone
respiratory complex II or succinate dehydrogenase - catalyzes transfer of e- from
succinate to ubiquinone
respiratory complex III or cytochrome b-c1 complex - catalyzes transfer of e- from
ubiquinone to cytochrome c
respiratory complex IV or cytochrome oxidase complex - catalyzes transfer of e-
from cytochrome c to oxygen

How is electron transfer coupled to ATP synthesis?

Process is known as oxidative phosphoryation
Respiratory complexes I, III, and IV yield enough energy for synthesis of 1 molecule of
ATP/ pair of electrons transferred --> act as coupling sites
Electrons from NADH encounter 3 coupling sites for ATP synthesis -->
Electrons from FADH2 encounter 3 complexes, but only 2 coupling sites -->

How is ATP synthesis actually carried out?

Proposed in 1961 by Peter Mitchell
Known as chemiosmotic coupling
energy released during electron transfer is used to pump protons from matrix into
intermembrane space
Establishment of an electrochemical proton gradient --> energy released when protons
move down electrochemical gradient through F1-F0 particles (ATP synthase) on inner
mitochondrial membrane

Eight lines of evidence upon which theory rests:

Electron transfer through respiratory chain causes protons to be pumped out of matrix
Thermodynamic calculations indicate proton gradient stores enough energy to drive ATP
formation
Artifically created pH gradients can drive ATP synthesis in absence of electron transport
Reconstituted membrane vesicles containing complexes I, III, or IV can establish proton
gradients
Uncoupling agents abolish the proton gradient and oxidative phosphorylation
dinitrophenol - inhibits oxidative. phosphorylation, but not electron transfer --> makes
membranes permeable to protons
carbonyl cyanide m-chlorophenylhydrazine (CCCP) - proton ionophore
gramicidin - same
valinomycin - K+ ionophore
nigericin - neutral exchange of H+ for K+ --> membrane potential unaffected, but pH
gradient abolished; oxidative phosphorylation requires intact membrane
Components of respiratory chain asymmetrically oriented within inner mitochondrial
membrane
ATP is synthesized as protons flow through F1-F0 complexes

3
Net equation for aerobic glucose oxidation:

C6H12O6 + 6 O2 ---> 6 CO2 + 6 H20

Fate of NADH molecules in cytosol

Not able to cross inner mitochondrial membrane
Must transfer its electrons to molecules that can pass so that these molecules can pass on
their electrons to respiratory chain
Two shuttle systems at work:
malate-aspartate shuttle - delivers electrons from cytoplasmic NADH to mitochondrial
NAD+.
glycerol phosphate shuttle - delivers electrons from cytoplasmic NADH to
mitochondrial FAD

Transport of other metabolites across inner mitochondrial membrane

Done through the use of carrier proteins
H+-linked pyruvate carrier - transports pyruvate into matrix with an inward flow of
protons
H -linked phosphate carrier - transports Pi into matrix with inward movement of
+

protons
ADP-ATP carrier - transports ADP into matrix and ATP into cytosol
Ca2+-H+ transporter - Ca2+ ions moved into matrix with exchange of protons
These processes often occur at the expense of ATP hydrolysis reduces the yield of
aerobic respiration.

Mitochondrial Myopathies

Term given to a group of neuromuscular diseases caused by damage to mitochondria DNA

(mtDNA)
mtDNA contains 13 structural genes that are all components of the respiratory chain
complexes

Symptoms
Depends upon which gene is affected.
Symptoms include:
Muscle weakness or exercise intolerance
Heart failure
Heart rhythm disturbances
Movement disorders
Stroke-like episodes
During exercise, muscles become easily fatigued or weak
Will have greatest effect on cells or organ systems with the highest energy requirements:
Brain

4
 Skeletal muscle
Cardiac muscle
Sensory organs
Kidneys

Treatment
Supportive only
Can take vitamin supplements such as:
Riboflavin
Coenzyme Q
Vitamin C and K
Carnitine

Other Topics of Interest:

Ethanol Intoxication
Caused by overdose of ethanol
Catabolism of ethanol depends upon amount of alcohol dehydrogenase in the liver
Causes decrease in NAD+ concentration.
Why?
Alcohol catabolism is catalyzed by two enzymes:
Alcohol dehydrogenase
Ethanol + NAD+  acetaldehyde + NADH +H+
Aldehyde dehydrogenase
Acetaldehyde + NAD+  acetate +NADH + H+
Effect is a dramatic increase in NADH levels .
Decreases aerobic utilization of glucose by inhibiting citric acid cycle
Remember those enzymes regulated by NADH levels?
Acetate is converted to acetyl CoA (precursor to fatty acid synthesis)

Methanol Toxicity
Is toxic to the liver because it is metabolized into formaldehyde by alcohol dehydrogenase
Effects:
Blindness due to damage to the optic nerve
Headache
Drowsiness
Seizures due to damage to brain tissue
Treatment involves administration of ethanol
Prevents metabolism of methanol to formaldehyde
Drug treatment with Antizol – competitive inhibitor of alcohol dehydrogenase, but
without the sedative effects of ethanol

5
Arsenic Poisoning
Metal often in trivalent form (AsO2-) as arsenite
Arsenate (pentavalent form; HAsO4-) substitutes for P in biological reactions  formation
of arsenate esters that are unstable

Effects of Arsenic Poisoning

Arsenite forms a stable complex with enzyme-bound lipoic acid
Causes inhibition of those enzymes that require lipoic acid as a coenzyme
Pyruvate dehydrogenase
α -ketoglutarate dehydrogenase

Brown Fat and Thermogenin

Brown fat is fat in newborns and hibernating animals that is used to help these organisms
thermoregulate
Example of an uncoupler (proteins that are transporters and disperse the proton gradient)
Burn brown fat (free fatty acids once broken down by b-oxidation) to generate an electron
flow
Electrochemical energy from electron transport is released as heat rather than ATP

Carbon Monoxide
Gas released from the combustion of almost any material
Competes with oxygen for binding sites on the hemoglobin molecule  reduces oxygen
transport to blood and produces hypoxia
CO has a higher affinity for Hb than does O2
Chemoreceptors in blood vessels are not triggered because CO does not significantly
decrease and amount of oxygen at lethal doses
Consequently, respiration is not increased to match the decrease in the blood’s ability to
carry oxygen.
Hypoxia slowly develops
Blood pressure drops because cardiac output is insufficient
Blood pools in peripheral tissues  fainting is a possibility
Other symptoms include headache, weakness, nausea, dizziness
Cyanide (CN) Poisoning

Inhibits cytochrome oxidase  halts electron transport, oxidative phosphorylation, aerobic

glucose metabolism
Results in lactic acidosis
Lethal dose is about 100 mg
Death usually occurs within an hour after ingestion