CHEM 450 - Biochemistry

Pyruvate
Dehydrogenase
&
Citric Acid Cycle
Dr. Felicia Ikolo, Ph.D., FIBMS, UK.
Department of Biochemistry, SGU
Email: fikolo@sgu.edu Tel. ext #: 3425
Office hours: 10.00 a.m. 11.30 a.m. M & W or
by appointment at other times
 Pyruvate Dehydrogenase & Citric Acid Cycle
Indicate how pyruvate is transported from the cytosol into the mitochondrial
matrix
Summarise the overall reaction catalysed by the PDH complex.
Outline the multienzyme nature of PDH complex and the cofactors that it
requires (TPP, lipoic acid, FAD)
Outline the regulation of the PDH complex (role of NADH, acetyl-CoA and
phosphorylation of PDH complex)
Indicate the inhibitory effects of trivalent arsenic compounds on PDH
Discuss the consequences of thiamine deficiency (Wernicke-Korsakoff
syndrome,Beri-Beri), and its effect on PDH and -ketoglutarate dehydrogenase
complexes.
Summarize the congenital abnormalities affecting the action of the PDH
complex and their clinical consequences.
Explain the central role of the TCA cycle in metabolism, including both its
catabolic and anabolic functions.
Outline the intermediates and the enzymes of the TCA cycle and CO 2
producing steps
Explain the energetics of the cycle, including reactions where reducing
equivalents (NADH and FADH2) are produced, and the production of GTP by
substrate level phosphorylation.
Catabolism of
proteins, fats, and
1: oxidation of fatty acids,
carbohydrates in the
glucose, and some amino
three stages of
acids yields acetyl-CoA.
cellular respiration.

2: oxidation of acetyl
groups in the citric acid
cycle includes four steps in
which electrons are
abstracted

3. electrons carried by
NADH and FADH2 are
funneled into the respiratory
chain and the production of
ATP.

3
 Important
Reactions of
Intermediary
Metabolism
 Alternate Fates of Pyruvate
We must link glycolysis to TCA Glucose
Glycolysis occurs in the
cytoplasm Glycolysis
End product of glycolysis is pyruvate Ace Ethanol
tal
Pyruvate is oxidized to acetyl-CoA de In yeast
hy
de and some
in mitochondrial matrix.
bacteria
Pyruvate
Reduction
Transamination

Oxidative
CarboxylationDecarboxylation
Alanine Lactate

Oxaloacetate Acetyl CoA

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PDH complex and TCA Cycle
PDH and TCA Cycle occur in the Matrix of Mitochondria.
Pyruvate
translocase

Pyruvate translocase
transports pyruvate into the
mitochondrial matrix
Pyruvate Dehydrogenase Complex
Complex of three enzymes:
Pyruvate dehydrogenase E1
dihydrolipoyl transacetylase E2
dihydrolipoyl dehydrogenase E3

Also contains two regulatory proteins:

a protein kinase and
a protein phosphatase
Requires 5 coenzymes:
Coenzyme A (CoA) derived fron vitamin B5
(pantothenic acid)
Flavin Adenine Dinucleotide (FAD) - from riboflavin
(vitamin B2)
Nicotinamide adenine dinucleotide (NAD+)-from
PDHC enzymes & their co enzymes
Enzyme Abbreviation Coenzyme
Pyruvate E1 TPP
dehydrogenas
e

Dihydrolipoyl E2 Lipioc acid &

transacetylase CoA

Dihydrolipoyl E3 FAD & NAD+

dehydrogenas
e

"Tender Loving care for Nancy"

Mechanism of Action of PDH Complex

L = lipoic acid
(C8H14O2S2)
 Regulation of PDH

Inhibition Activation
Dephosphorylati
Phosphorylation on
Insulin in
Acetyl CoA adipocytes and
ATP liver
NADH Catecholamines
in cardiac muscle
Calcium in
skeletal muscle

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NADH
PDH (inactive)
ATP
Acetyl-CoA
P P

PDH kinase
Regulation of PDH PDH phosphatas

+
PDH (active) Ca+2
Mg+2

Pyruvate + CoASH + NAD + Acetyl-CoA + NADH +CO2

 Deficiency of PDH Complex

iciency of the -subunit of E1 component of PDH complex (ra

ults in inability to convert pyruvate to acetyl CoA
Deficiency of PDH Complex
Pyruvate shunted to lactate via
LDH
Most common cause of
congenital lactic acidosis
Affects particularly the brain
which relies on TCA cycle for
most of its energy & most
sensitive to acidosis
Results in neurodegeneration,
muscle spasticity and early death
in neonatal onset form
X-linked dominant deficiency
Dietary restriction of
carbohydrates and
 Consequences of Thiamine Deficiency

PDH complex requires 5 coenzymes, including TPP

Wernicke syndrome & Korsakoff syndrome: related
disorders due to a deficiency of thiamine (vit. B1). The
term Wernicke-Korsakoff syndrome is used when the
two syndromes occur together usually in alcoholics and
malnourished individuals.
Wernicke's syndrome (Wernicke encephalopathy): neurological
disease characterized by confusion, the inability to coordinate
voluntary movement (ataxia), and eye (ocular) abnormalities.
Korsakoff's syndrome : mental disorder characterized by memory
loss
ffect of Arsenic compounds on PDH

Arsenic poisoning: due to its binding to lipoic acid which

is needed for the activity of dihydrolipoyl transacetylase
(E2)
Results in lactic acidosis, neurological defects especially
brain, can lead to death
Arsenic also affects other lipoic acid requiring enzymes
 Other Sources of Acetyl CoA

Glycogen Triglycerides Protein

glycogenolysis lipolysis proteolysis

glucose free fatty acids amino acids

glycolysis
oxidative
pyruvate oxidation deamination

oxidation

Acetyl CoA
Fates of Acetyl
CoA
TCA Cycle

Tricarboxylic acid
cycle
Krebs cycle
Citric Acid Cycle

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