Triacylcaboxylic Acid Cycle

Overview
• TCA cycle also called the Krebs cycle or the citric acid cycle, plays several roles in
metabolism.
• It is final pathway where the oxidative metabolism of carbohydrates, amino acids, &
fatty acids converge, their carbon skeletons being converted to CO2.
• This oxidation provides energy for the production of the majority of ATP in most
animals, including humans.
• The cycle totally occurs totally in the mitochondria & is, therefore, in close proximity to
the reactions of electron transport, which oxidize the reduced coenzymes produced by
the cycle.
• The TCA cycle is a an aerobic pathway, because O2 is required as the final electron
acceptor.
• The TCA cycle also participates in a number of important synthetic reaction e.g
formation of glucose from the carbon skeletons of some amino acids, and it provides
building blocks for the synthesis of some amino acid and heme.
• The cycle should not be viewed as a closed circle, but instead as a traffic circle with
compounds entering & leaving as required.

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 The Citric Acid Cycle
The citric acid cycle is the final common pathway for the oxidation of fuel molecules:
amino acids, fatty acids, & carbohydrates.

• Most fuel molecules enter the cycle as acetyl coenzyme A

• This cycle is the central metabolic hub of the cell
• It is the gateway to aerobic metabolism for any molecule that can be transformed
into an acetyl group or dicarboxylic acid,
• It is also an important source of precursors for building blocks
• Also known as, Krebs Cycle, & Tricarboxylic Acid Cycle (TCA)

ØThe citric acid cycle oxidizes two-carbon units

ØEntry to the cycle and metabolism through it are controlled
ØThe cycle is a source of biosynthetic precursors

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 Introduction
1. The function of the cycle is the harvesting of high-energy
electrons from carbon fuels
2. The cycle itself neither generates ATP nor includes O2 as a
reactant
3. Instead, it removes electrons from acetyl CoA & uses them to
form NADH & FADH2 (high-energy electron carriers)
4. In oxidative phosphorylation, electrons from reoxidation of
NADH & FADH2 flow through a series of membrane proteins
(electron transport chain) to generate a proton gradient
5. These protons then flow back through ATP synthase to
generate ATP from ADP & inorganic phosphate
6. O2 is the final electron acceptor at the end of the electron
transport chain
7. The cytric acid cycle + oxidative phosphorylation provide
> 95% of energy used in human aerobic cells

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Citric Acid Cycle: Overview
Input: 2-carbon units
Output: 2 CO2, 1 GTP,
& 8 high-energy
electrons

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Dehydration synthesis or condensation reaction

Oxaloacetate 4 C molecule:
Acetyl CoA transfers its acetyl group to
the #2 carbonyl carbon via
the methyl end.
This forms a 6 C citrate molecule.
 First dehydration then hydration. Why?
Aconitase enzyme catalyzes both reactions

Citrate:
Note that it is a tertiary alcohol which is not oxidizable.
Isomer is a 2o alcohol (isocitrate) that can be oxidized
 First of 4 Redox reactions,
followed by decarboxylation.
NAD+ is the oxidizing agent.
Isocitrate dehydrogenase is the catalyst

Alpha ketoglutarate (a ketone) is formed

when Isocitrate is oxidized,
leading to the reduction of NAD+ to NADH,
& also decarboxylated (first CO2 formed)
 Redox reaction #2:
Oxidation of a-ketoglutarate
into 4 C Succinyl CoA (also a ketone)
again more CO2 and NADH are formed

Three enzymes used as a group:

a-ketoglutarate dehydrogenase complex.
H atom must be removed from
sulfhydryl group for coenzyme A to form
a high energy thioester bond.
 Pyurvate dehydrogenase complex
A large, highly integrated complex of three kinds of enzymes

Pyruvate + CoA + NAD+ Þ acetyl CoA + CO2 + NADH

Groups travel from one active site to another, connected by

tethers to the core of the structure

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Summary of total energy yield of complete
oxidation of 1 glucose molecule
Step Coenzyme ATP Source of ATP
Yield Yield
Glycolysis –Stage 1 - 2 Phosphorylation of glucose and
fructose uses 2 ATP
Glycolysis –Stage 2 4 Substrate level phosphorylation
2 NADH 6 Oxidative phosphorylation
Pyruvate metabolism 2 NADH 6 Oxidative phosphorylation
TCA cycle 2 Substrate level phosphorylation
6 NADH 18 Oxidative phosphorylation
2 FADH2 4 Oxidative phosphorylation
Total Yield 38 ATP
 Inhibitors of TCA Cycle
• Fluoroacetyl CoA:
-It inhibits aconitase enzyme
-It combines with oxaloacetate giving rise to fluorocitrate .
• Malonic acid:
-Inhibits succinate dehydrogenase (competitive
inhibition)
• Arsenate and Mercury :
-Inhibit Pyruvate dehydrogenase and α-ketoglutarate
dehydrogenase complexs.
- By reacting with sulphydral group of lipoic acid leading
to accumulation of pyruvic lactic acid and α-
ketoglutarate.
Regulation of CAC:
Rate controlling enzymes:
Citrate synthatase
Isocitrate dehydrogenase
a-keoglutaratedehydrogenase

Regulation of activity by:

Substrate availability
Product inhibition
Allosteric inhibition or activation by
other intermediates

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 Regulation of pyruvate dehydrogenase

Inhibited by products,
NADH & Acetyl CoA

Also regulated by covalent modification,

the kinase & phosphatase also regulated

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Biosynthetic roles of the citric acid cycle

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Arsenic Compound poisoning: Inactivation of E-2 of PDC, and other proteins.

Organic Arsenical were used

as antibiotics for the treatment
of syphilis and
trypanosomiasis.
Micro-organisms are more
sensitive to organic arsenicals
than humans.
But these compounds had
severe side effects and As-
poisoning.
Fowler’s solution, the famous
19th century tonic contained
10mg/ml As. Charles Darwin
died of As poisoning by taking
this tonic.
Napoleon Bonaparte’s death
was also suspected to be due
to As poisoning.

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Thiamin (Vitamine B1) deficiency causes Beriberi:
Thiamine pyrophosphate (TPP) is an important cofactor of pyruvate
dehydrogenase complex, or PDC a critical enzyme in glucose metabolism.
Thiamine is neither synthesized nor stored in good amounts by most vertebrates.
It is required in the diets of most vertebrates. Thiamine deficiency ultimately
causes a fatal disease called Beriberi characterized by neurological disturbances,
paralysis, atrophy of limbs and cardiac failure. Note that brain exclusively uses
aerobic glucose catabolism for energy and PDC is very critical for aerobic
catabolism. Therefore thiamine deficiency causes severe neurological symptoms.
Arsenic Poisoning: Arsenic compounds such as arsenite (AsO3---) organic
arsenicals are poisonous because they covalently bind to sulfhydryl compounds
(SH- groups of proteins and cofactors). Dihydrolipoamide is a critical cofactor of
PDC, and it has two-SH groups, which are important for the PDC reaction. These
–SH groups are covalently inactivated by arsenic compounds as shown below;
OH HS S
-O As + -O As + 2H2O
OH HS S
R R

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Arsenic compounds in low doses are very toxic to microorganisms,
therefore these compounds were used for the treatment of syphilis and
other diseases in earlier days. Arsenicals were first antibiotics, but with a
terrible side effects as they are eventually very toxic to humans.
Unfortunately and ignorantly, a common nineteenth century tonic, the
Fowler’s solution contained 10 mg/ml arsenite. This tonic must have
been responsible for many deaths, including the death of the famous
evolution scientist Charlse Darwin.

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