BIOCHEMISTRY

2.03 PYRUVATE DEHYDROGENASE COMPLEX AND THE

TRICARBOXYLIC ACID CYCLE LE2
Dr. Reyes | September 27, 2016

I.
TRI
OUTLINE

Pyruvate Dehydrogenase Complex

#
A. Pyruvate Dehydrogenase (E1)
B. Dihydrolipoyl Transacetylase (E2)
C. Dihydrolipoyl Dehydrogenase (E3)
II. The Tricarboxylic Acid Cycle
A. Introduction to the TCA Cycle
B. Reactions of the TCA Cycle
1. Synthesis of Citrate from Acetyl CoA
and Oxaloacetate
2. Isomerization of Citrate to Isocitrate
3. Oxidation and Decarboxylation of
Isocitrate
4. Oxidation and Decarboxylation of α-
ketoglutarate
5. Cleavage of Succinyl CoA
6. Oxidation of Succinate to Fumarate
7. Hydration of Fumarate Figure 1. The Pyruvate Dehydrogenase Complex.
8. Oxidation of Malate to Oxaloacetate
C. Shuttle Mechanism for Cytoplasmic  Once inside the mitochondrion, Pyruvate is oxidatively
Reducing Equivalents decarboxylated to Acetyl CoA by pyruvate
D. ATP Yield for Complete Oxidation of
dehydrogenase complex
Glucose
E. Amphibolic Nature of the TCA Cycle  Composed of 3 enzymes and 5 coenzymes
F. Regulation of the TCA Cycle  Made up of 5 steps
G. Role of Vitamins in the TCA Cycle  The overall reaction is exergonic and irreversible; the
H. Anaplerotic Reactions of the TCA Cycle ΔG of the reaction is -8.0 kcal/mole
I. Catabolism of Major Nutrients and Their
Convergence with the TCA Cycle
J. Clinical Correlations of the TCA Cycle Note: Exergonic reactions are usually irreversible
1. Beri-beri
2. Chronic Alcoholism
 The three enzymes/coenzymes comprising the pyruvate
III. Mini Quiz
IV. Tables, Images and Figures dehydrogenase complex are as follows:

OBJECTIVES: Table 2. Enzyme components of PDH complex.

At the end of the lecture, the student should be able to: ENZYME CODE COENZYME REACTION
1. Describe the overall process of the TCA cycle and PDH Oxidative
Pyruvate
complex. E1 TPP decarboxylation
dehydrogenase
2. Determine the substrates, enzymes, products, and of pyruvate
amount of energy utilized and produced in the two Dihydrolipoyl Transfer acetyl
E2 Lipoamide
pathways. transacetylase group to CoA
3. Discuss the mechanisms by which the pathways are Regenerates
Dihydrolipoyl
regulated E3 FAD oxidized
dehydrogenase
lipoamide
Legend:
Remember Previous  Other coenzymes of pyruvate dehydrogenase:
Lecturer Book Trans Com
(Exams) Trans o Coenzyme A (CoA)
     o Nicotinamide adenine dinucleotide (NAD+)
 Congenital Lactic Acidosis: deficiency in pyruvate
I. PYRUVATE DEHYDROGENASE (PDH) COMPLEX dehydrogenase  inability to convert pyruvate to acetyl
CoA  pyruvate converted to lactic acid by lactate
 Pyruvate, a 3C compound, is a product of glycolysis; it dehydrogenase  may lead to brain damage
enters the mitochondrion via pyruvate translocase o The brain uses pyruvate oxidation and the TCA
and in the presence of CoASH (Coenzyme A). cycle as energy source
 The PDH complex is responsible for converting
pyruvate to CO2 and the acetyl potion of acetyl-coA

 NOT part of the TCA cycle proper links  the glycolytic
pathway with the TCA
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A. Pyruvate Dehydrogenase (E1) B. Dihydrolipoyl Transacetylase (E2)

STEP 1: STEP 3:

Figure 4. Step 3 of the Pyruvate Dehydrogenase Complex.

 CoA-SH reacts with the lipoamide-bound acetyl group

Figure 2. Step 1 of the Pyruvate Dehydrogenase Complex. of E2, forming acetyl CoA and leaving the lipoamide in
the reduced dithiol (SH-SH) form
 Pyruvate reacts with thiamine pyrophosphate (TPP,
prosthetic group of E1) via pyruvate dehydrogenase C. Dihydrolipoyl Dehydrogenase (E3)
 CO2 is released and hydroxyethylthiamine
pyrophosphate (HETPP) is formed STEP 4:
Note: Decarboxylation means release of carbon dioxide

STEP 2:

Figure 5. Step 4 of the Pyruvate Dehydrogenase Complex.

 The prosthetic group of E3-FAD oxidizes the reduced

lipoamide of E2, forming FADH2
 This re-oxidation allows E2 to participate in multiple
rounds of the reaction

STEP 5:

Figure 6. Step 5 of the Pyruvate Dehydrogenase Complex.

Figure 3. Step 2 of the Pyruvate Dehydrogenase Complex.
 NAD+ is reduced by E3-FADH2 to regenerate E3-FAD
 The 2-carbon hydroxyethyl fragment is transferred to  The oxidation of E3-FADH2 regenerates the original
holoenzyme, completing the catalytic cycle
the lipoamide group of E2 catalyzed by E1
 NADH dissociates into the mitochondrial matrix where
 This prosthetic group consists of lipoic acid covalently it serves as a mobile carrier of reducing energy
bound by amide linkage to a lysine residue in E2
 The oxidation of HETPP is coupled to the reduction of
the disulfide of lipoamide
 The acetyl group is then transferred to a sulfhydryl
group of dihydrolipoamide

Note: Lipoamide has a disulfide group which makes it

easily reducible

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II. THE TRICARBOXYLIC ACID (TCA) CYCLE B. Reactions of the TCA Cycle

A. Introduction to the TCA Cycle 1. Synthesis of Citrate from Acetyl CoA and
Oxaloacetate

Figure 7. Overview of glycolysis and TCA cycle.

Figure 9. Synthesis of citrate.
 Other names: Krebs cycle, Tricarboxylic Acid Cycle
 Site: Mitochondrial matrix  Reaction: Irreversible, Exergonic
 Enzyme: Citrate Synthase
Identical intermediates can be used for different  Substrate: Acetyl CoA
purposes inside and outside the mitochondria. The main fate of  Product: Citrate
mitochondrial acetyl-coA is oxidation in the TCA cycle (in the  Methyl carbon of acetyl CoA + carbonyl carbon of
cytoplasm, it is used for biosynthesis of fatty acids and oxaloacetate
cholesterol)  Only reaction with C-C bond formation
Major function: The final common pathway for the  Rate limiting
oxidation of all fuels (fat, carbohydrates, proteins, and lipids)
via the formation of a common metabolite Acetyl CoA 2. Isomerization of Citrate to Isocitrate
Two major functions: energy production and
biosynthesis; End products: CO2, 3 NADH/ FADH2, ATP

Figure 10. Isomerization of citrate.

 Reaction: Isomerization of Citrate (Endergonic,

Reversible)
 Enzyme: Aconitase/ Aconitase Hydratase
 Citrate cannot be oxidized directly, therefore, is
isomerized to Isocitrate
o The rearrangement of citrate to Isocitrate is
Figure 8. Overview of the TCA cycle. done by moving the hydroxyl group
o The hydroxyl group is not in a desirable
 Acetyl CoA is not part of the cycle. It is converted to place to be acted upon by the rest of the
citrate in the TCA cycle. dehydrogenase enzymes, so it needs to be
isomerized to an isocitrate.
No oxygen is used in any of the reactions. The cycle  Intermediate formed: Cis-Aconitate
requires oxidative metabolism in the mitochondrion for re- o Isomerized to enable isocitrate to undergo
oxidation of reduced coenzymes. Reduced coenzymes provide oxidative decarboxylation via Cis-Aconitate.
energy for ATP synthesis by the electron transport system. o The aconitase enzyme dehydrates first
Defects in the TCA cycle lead to limited ATP production. Cells (taking away of H2O) then hydrates again
then die rapidly or are severely impaired functionally. Tissues (insertion of H2O)
that use oxygen at rapid rates: CNS, muscles

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 2.03 PDH COMPLEX AND THE TCA CYCLE BIOCHEMISTRY 2020C

o Aconitase is an example of a moonlighting

enzyme which is an enzyme that it is
capable of performing multiple functions.
 closely resembles the iron receptor
protein (IRP)
 could also help recognize anemia and
actually help the body cover for that
anemia, so that it could recover while
waiting for the iron stores to be
recovered by the body.
o Although the aconitase reaction does not
require cofactors, it requires ferrous (Fe 2+)
iron in its catalytic mechanism. This Fe 2+ is
involved in an iron–sulfur center, which is an
essential component in the hydratase activity
of aconitase (Devlin, 1997).
 Inhibition: Fluorocitrate is a potent inhibitor of Figure 12. Oxidation and decarboxylation of α-ketoglutarate.
aconitase. It comes from the condensation of
fluoroacetyl-CoA and oxaloacetate  α-ketoglutarate dehydrogenase complex has a
similar structure to Pyruvate Dehydrogenase
3. Oxidation and Decarboxylation of Isocitrate (PDH)
 NAD+ is reduced to NADH by accepting an H
atom from the carboxyl group of α-ketoglutarate
and another H atom from CoASH.
 CO2 comes from the cleavage of one of the
carboxyl groups of α-ketoglutarate.
 This reaction is a key regulatory step of the Krebs
cycle.
 Inhibition: Arsenite inhibits α-ketoglutarate
dehydrogenase

5. Cleavage of Succinyl CoA

Figure 11. Oxidation and decarboxylation of isocitrate.

 Reaction: Oxidation and decarboxylation of

Isocitrate (Exergonic, Irreversible)
 Enzyme: Isocitrate dehydrogenase (key or major
regulatory enzyme of the Krebs cycle)
 First Oxidative Carboxylation (or Dehydrogenase
Reaction) in TCA
 Isocitrate is oxidatively decarboxylated to α-
ketoglutarate; producing the 1st of the 3 NADH of
the cycle and releasing the 1st CO2 of the cycle.
o Oxalosuccinate is decarboxylated. The beta
carboxyl group of oxalosuccinate is cleaved
to form α-ketoglutarate.
 Oxalosuccinate is an unstable keto acid
intermediate form that is bound to Isocitrate
Dehydrogenase.
 This reaction is one of the control points of the
cycle
4. Oxidation and Decarboxylation of α-ketoglutarate
 Reaction: Oxidative decarboxylation of α-
Ketoglutarate (Highly Exergonic, Irreversible)
 Enzyme: α-ketoglutarate dehydrogenase complex
 Second Oxidative Decarboxylation in TCA
 α-ketoglutarate is oxidatively decarboxylated by α-
ketoglutarate dehydrogenase complex to succinyl
CoA; producing the 2nd NADH and releasing the
2nd CO2 of the cycle.
Figure 13. Cleavage of succinyl CoA.
 Reaction: Exergonic, Irreversible
 Enzyme: Succinyl CoA synthetase/ Succinyl
thiokinase
 Succinyl CoA synthetase (succinate thiokinase)
catalyzes the conversion of energy-rich succinyl-
CoA to succinate and free CoA
 The high energy thioester bond in succinyl-CoA is
used to generate GTP from GDP and Pi in the
reaction (Mg2+ aids transfer of PO4 to GDP)
 This is the only substrate-level phosphorylation
reaction in the Citric Acid Cycle
o Substrate-level phosphorylation – high-
energy thioester bond serves as the driving
force for the synthesis of a high-energy
phosphate bond (in this case, GTP)
 GTP formed provides a regulatory link between
citric acid cycle activity and the withdrawal of

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oxaloacetate for gluconeogenesis (link between

gluconeogenesis & TCA cycle). 7. Hydration of Fumarate

Note: GTP = 1 mol ATP

 Kidney and liver tissues (gluconeogenesis sites)
have 2 isoenzymes for succinyl thiokinase (1ATP,
1GTP)
 Non-gluconeogenic tissue have only 1 type (ATP)

6. Oxidation of Succinate to Fumarate

Figure 15. Hydration of fumarate.

 Enzyme: Fumarase/ Fumarate hydratase

 Reaction: Endergonic, Reversible
 Fumarase/ fumarate hydratase is a tetramer and is
highly stereospecific for the trans conformation of
the substrate (the cis form, maleate, is NOT a
substrate).
 This enzyme stereospecifically adds water across
the trans double bond (between C2 and C3) of
fumarate to form α-hydroxy acid, L-malate.

8. Oxidation of Malate to Oxaloacetate

Figure 14. Oxidation of succinate.

 Enzyme: Succinate dehydrogenase/ Complex II

 Reaction: Endergonic, Reversible
 Succinate dehydrogenase, unlike any other
enzymes in TCA that can be found in the
mitochondrial matrix, is tightly bound to the inner
mitochondrial membrane where it is a part of
Complex II (succinate-Q reductase)
 reason why it is alternatively called as Complex
II

Note: Succinate dehydrogenase is the only enzyme of the

TCA that is not found in the mitochondrial matrix Figure 16. Oxidation of malate

 Two subunits of succinate dehydrogenase  Enzyme: Malate dehydrogenase

o 70,000-MW (molecular weight) subunit  Reaction: Endergonic, Reversible
contains the substrate-binding site,  Malate is oxidized to oxaloacetate by malate
covalently-bound FAD (to a lysine residue), 4 dehydrogenase; producing the (3rd and final
nonheme iron atoms, and 4 acid-labile sulfur NADH) of the cycle.
atoms  The alcohol group of malate is oxidized to a keto
o 30,000-MW (Iron-sulfur protein); contains 4 group through the donation of electrons to
nonheme irons, and 4 acid labile sulfur NAD+
atoms  The conversion of malate to oxaloacetate
completes one round trip through the TCA cycle.
 This is the first dehydrogenation reaction. Succinate is  The oxaloacetate may then react with acetyl-
oxidized to the trans-dicarboxylic acid fumarate, with reduction CoA to continue another cycle of reactions.
of FAD to FADH2. Single electrons are transferred from the two  This reaction is analogous to the lactate
adjacent -CH2- methylene groups of succinate to a FAD bound dehydrogenase reaction in glycolysis.
to succinate dehydrogenase, thereby forming the double bond
of fumarate. Malonate is a competitive inhibitor of succinate
dehydrogenase wherein the dicarboxylic acid binds the active
Summary:
site of succinate dehydrogenase but doesn’t undergo oxidation
The Citric Acid Cycle is the final pathway for the oxidation of
because of a lack of –CH2-CH2 bond.
carbohydrates, lipids, and proteins. Their common end
metabolite is Acetyl CoA, which reacts with oxaloacetate to
form citrate. Regeneration of oxaloacetate is done by a series

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 2.03 PDH COMPLEX AND THE TCA CYCLE BIOCHEMISTRY 2020C

of oxidative reduction, decarboxylation and hydration with the Table 2. ATP yield for glycolysis.
help of reducing enzymes and coenzymes that release NAD
and CO2. Reaction Mechanism ATP Yield

TCA cycle is a sequence of eight enzymatic reactions, two Isocitrate NADH, Oxidative
5
of which involve decarboxylations. One molecule of GDP is dehydrogenase Phosphorylation
phosphorylated to GTP; one molecule of FAD is reduced to
FADH2; and four molecules of NADH are produced (3 from the α-ketoglutarate NADH, Oxidative
5
citric acid cycle, 1 from the reaction of the pyruvate dehydrogenase Phosphorylation
dehydrogenase complex).
Succinyl CoA Substrate Level
2
C. Shuttle Mechanism for Cytoplasmic Reducing synthase Phosphorylation
Equivalents
Succinate FADH2, Oxidative
 NADH are also sources of energy upon reoxidation to 3
dehydrogenase Phosphorylation
NAD+ in the electron transport chain
 However, they are impermeable to the mitochondrial
membrane so they use shuttles (malate-aspartate and Malate NADH, Oxidative
5
glycerol-phosphate shuttles) to be transported in the dehydrogenase Phosphorylation
inner mitochondrial membrane
1. The Malate-aspartate shuttle makes use of the fact
that malate can cross the mitochondrial membrane, Table 3. ATP yield for glycolysis and TCA cycle.
while oxaloacetate cannot. Mol
o Electron transfer from NADH in the cytosol
Reaction Mechanism ATP/Mol
produces NADH in the mitochondrion
Glucose
o Cytosolic malate dehydrogenase catalyzes
Hexokinase-
the reduction of oxaloacetate to malate; NADH Phosphorylation -1
Glucokinase
is also oxidized to NAD+ → malate crosses
GLYCOLYSIS

mitochondrial membrane PFK-1 Phosphorylation -1

o Mitochondrion malate dehydrogenase
catalyzes the conversion of malate to G-3-PDH NADH, Oxid. Phos +2.5 (1.5)
oxaloacetate; oxaloacetate is converted to Phosphoglce- Substrate-level
aspartate → aspartate crosses mitochondrial +2
rate kinase Phospho.
membrane Substrate-level
o Aspartate is converted to oxaloacetate in the Pyruvate kinase +2
Phospho.
cytosol, completing the cycle Pyruvate
o NADH produced in the mitochondrion passes NADH, Oxid. Phos +5
dehydrogenase
electrons to ETC
Isocitrate +5
2. The Glycerol-phosphate shuttle uses NADH and NADH, Oxid. Phos
dehydrogenase
produces FADH2 inside the cell.
α-ketoglutarate +5
o glycerol phosphate is produced by the reduction NADH, Oxid. Phos
TCA

dehydrogenase
of dihydroxyacetone phosphate; NADH is
oxidized to NAD+ Succinyl CoA +2
Substrate-level Phos
o FAD acts as the oxidizing agent and is reduced synthetase
to FADH2 → passes electrons to ETC Succinate +3
FADH2 Oxid. Phos
 Malate-aspartate shuttle = 2.5 ATP/NADH molecule dehydrogenase
 2 NADH x 2.5 = 5 ATP Malate +5
NADH Oxid. Phos.
 Glycerol-phosphate shuttle = 1.5 ATP/NADH2 dehydrogenase
molecule TOTAL 30-32
 2 NADH2 x 1.5 = 3 ATP
Overall reaction:
3. ATP Yield for Complete Oxidation of Glucose Acetyl CoA + 3 NAD+ + 1 GDP + Pi + 2 H2O  CoA-SH + 3
NADH + 3 H+ + 1 GTP + 2 CO2

 For every Acetyl CoA:

4. Amphibolic Nature of the TCA Cycle
o 1 NADH = 2.5 ATPs
o 1 FADH = 1.5 ATPs  Can participate in either anabolic or catabolic reactions
 For every two Acetyl CoA:  Can act as a precursor for proteins, fatty acids and
o Total = 20 glucose synthesis
 Pyruvate  NADH, Oxidative Phosphorylation  5  The 4 carbon (Oxaloacetate) and 6 carbon (Citrate) are
ATP important intermediates in the cycle
 2 pyruvates oxidized through the TCA  32 ATPs  Amino Acid Synthesis: transamination reactions
(depends of tissue involved and shuttle used) o Pyruvate  production of alanine and other aa

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 2.03 PDH COMPLEX AND THE TCA CYCLE
o α-ketoglutarate  can become glutamate / GABA
or glutamine
o Oxaloacetate  transaminated to aspartate 
converted to urea, purines, pyrimidines, and
cytosine
Figure 17. Diagram showing the amphibolic nature of the TCA Cycle.
 Fatty Acid and Steroid Synthesis: occurs when
isocitrate dehydrogenase is inhibited by the
accumulation of ATP and NADH, leading to the export
of citrate to the cytosol and releasing Acetyl-CoA by
citrate lyase
 Heme Synthesis: Succinyl CoA couples with glycine
and if acted upon ALAS (ALA synthase) 
aminolevulinic acid is formed
 Gluconeogenesis: oxaloacetate can be converted to
malate, which can enter gluconeogenesis in times of
starvation
Figure 18. Diagram of the TCA Cycle showing the precursors to
the different intermediate compounds.
5. Regulation of the TCA Cycle
 The regulation and control of TCA cycle is chiefly
based on the level of the 5 enzymes that catalyze
irreversible reactions (non-equilibrium reactions):
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o Pyruvate dehydrogenase
o Citrate synthase
o Isocitrate dehydrogenase
o a-ketoglutarate dehydrogenase
o Malate dehydrogenase
 The main regulatory mechanism employed is by
allosteric control.
Figure 19. Diagram showing the regulation of the TCA Cycle.
NOTE: ATP, NADH and the products of the cycle
are actually inhibitory in dehydrogenases. ADP, NAD,
and Ca++ ions are stimulatory to the cycle.
When there is a high ATP: ADP ratio and a high NAD:
NADH ratio, there will be enough energy running
through the tissue or cell (example is muscle) and this
energy status will be the one to signal to your body that
it can stop producing more ATP/energy. Therefore, this
is your control/regulation. 
 Pyruvate dehydrogenase
o Allosteric inhibitors: ↑ ATP, ↑ NADH, ↑ Acetyl CoA
OR sufficient energy supply in the cell in its resting
state
o Allosteric activator: Ca++ ions
 Citrate synthase
o Allosteric inhibitors: ↑ ATP, ↑ NADH, ↑ Succinyl
CoA, long-chain fatty acyl CoA, citrate (product
inhibitor)
 Isocitrate dehydrogenase
o Allosteric inhibitors: ↑ ATP, ↑ NADH
o Allosteric activators: ADP, NAD+, Ca++ ions
(released by ER of muscle tissues and provide
additional activation during hydrolysis of ATP)
 a-ketoglutarate dehydrogenase
o Allosteric inhibitors: ↑ ATP, ↑ GTP, ↑ NADH, ↑
Succinyl CoA
o Allosteric activator: Ca++ ions

Note: Succinyl CoA regulates its own synthesis making α-
ketoglutarate available for other biosynthetic reactions.
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 2.03 PDH COMPLEX AND THE TCA CYCLE BIOCHEMISTRY 2020C

 Malate dehydrogenase o Other amino acids includes arginine, histidine,

o Allosteric inhibitors: ↑ATP, ↑ NADH glutamine, and proline.
 Synthesis of succinyl CoA from propionyl CoA:
6. Role of Vitamins in the TCA Cycle o Isoleucine, methionine, and valine also contributes
to the formation of succinyl-CoA
o The same goes with cholesterol and odd chain
Table 4. Role of vitamins in the TCA cycle.
fatty acids
ACTIVE
VITAMIN FUNCTION
FORM
Coenzyme for decarboxylation 8. Catabolism of Major Nutrients and Their Convergence
by the pyruvate with the TCA Cycle
Thiamine
dehydrogenase complex,
Thiamine Pyrophosphate
isocitrate dehydrogenase & α-
(TPP)
ketoglutarate dehydrogenase
complex (Catalytic cofactors)
Flavin Adenine Cofactor of succinate
Riboflavin Dinucleotide dehydrogenase (Catalytic
(FAD) cofactor)
Electron acceptor for isocitrate
Nicotinamide dehydrogenase, α-
Adenine ketoglutarate dehydrogenase
Niacin
Dinucleotide complex & malate
(NAD) dehydrogenase
(Stoichiometric cofactor)
Cofactor attached to "active"
Pantothenic Component of carboxylic acid residues, such
Figure 20. Interconnections of different metabolic reactions with the TCA
Acid Coenzyme A as Acetyl CoA and Succinyl
Cycle.
CoA (Stoichiometric cofactor)
 Acetyl CoA is the common product of protein,
carbohydrate and fat metabolism
7. Anaplerotic Reactions of the TCA Cycle
 Acetyl CoA enters the TCA cycle where GTP, NADH,
 Pathways or reactions that replenish the intermediates FADH2, and carbon dioxide is generated.
of the TCA cycle  “filling up” reactions  NADH and FADH2 is then transported to the ETC,
 Removal of any of the intermediates from the TCA which is used to drive the ATP synthase, to produce
cycle removes the four carbons used to regenerate ATP
Oxaloacetate during each turn of the cycle → depletion
of Oxaloacetate → impossible to continue oxidation of 9. Clinical Correlations of the TCA Cycle
Acetyl CoA.
a. Beri - Beri
 To keep the TCA cycle running, cells have to supply
 A neurologic and cardiovascular disorder stemming
enough 4-carbon intermediates from degradation of from impaired glucose oxidation caused by dietary
carbohydrate or certain amino acids to compensate for deficiency of thiamine (Vitamin B1)
the rate of removal (Anaplerotic Reactions)  Common among population with rice as the major
 Pyruvate carboxylase reaction: the major anaplerotic food (Far East).
reaction in the body (liver and kidney).  Aggravated by eating polished rice whose outer
o Catalyzed by pyruvate carboxylase that converts layer, rich in thiamine, is removed by excessive
washing before cooking.
pyruvate to oxaloacetate
 PATHOPHYSIOLOGY OF BERI-BERI: Deficiency
o Rate limiting step of gluconeogenesis of thiamine → ↓ TPP → impaired glucose oxidation
o Uses biotin as cofactor for transferring CO₂ to esp. in the nervous system → ↓ ATP: muscle
Oxaloacetate → Uses ATP (endergonic) weakness and atrophy, pain in the limbs, fatigue,
o Pyruvate can come from alanine, cysteine, cardiac enlargement, ↓ CO and edema due to ↓
glycine, hydroxyproline, serine, threonine, venous return
b. Chronic alcoholism
tryptophan
 Ethanol consumption leads to accumulation of
 Reversible conversion of glutamate to α-
NADH  blocked processing of acetyl CoA in TCA
ketoglutarate: the second major anaplerotic reaction in because ↑ NADH inhibits dehydrogenases
the body (isocitrate dehydrogenase and a-ketoglutarate
o Catalyzed by transaminase (TA) and glutamate dehydrogenase)
dehydrogenase (GDH)

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 Leads to excess Acetyl CoA  ketone bodies will

be formed and released in the blood  more acidic
internal environment along with lactate build-up and
accumulation of acetaldehyde in the liver 
impaired protein function leading to liver damage.

MINI QUIZ
1. Which of the following is not a Krebs cycle inhibitor?
a. Malonate
b. Arsenite
c. Malate
d. Fluorocitrate

For items 2-6: Match the enzyme and its product in the TCA
2. Succinate a. Fumarate
dehydrogenase
3. Aconitase b. Succinyl CoA
4. Fumarase c. Citrate
d. Acetyl CoA
5. Pyruvate e. Isocitrate
dehydrogenase
6. Succinyl Thiokinase f. Malate
g. Succinate

Answers: C A E F D G

REFERENCES:
1. Lecture Notes
2. Madarcos Notes
3. 2019A Transes
4. Baynes, John W., Medical Biochemistry, 4th ed.

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