TCA Cycle

Lippincott’s Illustrated Reviews: Biochemistry,

MOL 114

Dr Hanaa Hajeer
hhajeer@alfaisal.edu.sa

https://www.khanacademy.org/test-prep
/mcat/biomolecules/krebs-citric-acid-cycl
e-and-oxidative-phosphorylation/v/krebs-
citric-acid-cycle
Aerobic respiration

2
 The Pyruvate Dehydrogenase complex
• Pyruvate is shuttled into
mitochondria with the help of a
dedicated transporter.
Pyruvate
• The Pyruvate Dehydrogenase
complex (PDC) is a gigantic NAD+
PDC
multi-enzyme complex with
NADH+H+
dozens of copies each of three CO2
enzymes E1, E2 and E3. Acetyl-CoA

Oxaloacetate Citrate

Malate Isocitrate
CO2
Fumarate α-Ketoglutarate
CO2
Succinate
Succinyl-CoA
Pyruvate dehydrogenase
 Pyruvate dehydrogenase require 5 coenzymes
The Lovely Co-enzymes For Nerds
1. Thiamine pyrophosphate (B1)
2. Lipoic acid
3. CoA (B5, pantothenic acid)
4. FAD (B2, riboflavin)
5. NAD+ (B3, niacin)
 PDH is inhibited by arsenite
• Arsenic is odourless and tasteless heavy metal
• Used for centuries as poison
• Results in impaired production of acetyl CoA and of
subsequent energy production via oxidative
phosphorylation
• Arsenic occurs in two forms : Arsenate and arsenite
Arsenite inhibits lipoic acid
clinical findings: Mee lines
Genetic mutation: Pyruvate dehydrogenase deficiency
• Deficiency in one of component of pyruvate
dehydrogenase complex is most common biochemical
cause of congenital lactic acidosis
• Results in inability to convert pyruvate to acetyl CoA
• Causing pyruvate to be converted to lactic acid via
lactate dehydrogenase
• This causes problems for brain, which relies on the
TCA cycle for most of its energy, and is particularly
sensitive to acidosis
• Symptoms include hypotonia (weak muscle), lactic
acidosis, and seizures.

7
 The TCA cycle
• The TCA cycle (or Krebs cycle, or citric acid cycle) is a central “metabolic roundabout”
with multiple entry and exit points. Several of the intermediates are involved in
gluconeogenesis, amino acid and heme metabolism.

• The oxidative catabolism of carbohydrates, lipids and amino acids comes together
here.

• All TCA cycle reactions happen in mitochondria and require oxygen to recycle the
reduced coenzymes NADH+H+ and FADH2

Acetyl-CoA

Oxaloacetate Citrate

Malate Isocitrate
CO2
Fumarate α-Ketoglutarate
CO2
Succinate
Succinyl-CoA
 TCA cycle function
• Four intermediates of the TCA
• The TCA cycle provides for full
cycle are amino acid
oxidation of acetyl-CoA to 2 CO2
metabolites. This allows their
conversion to glucose by
• Oxidation of NADH and FADH2 Fatty acids gluconeogenesis.
in the mitochondria generates Amino acids
Carbohydrates
28 ATP per molecule glucose
and 2 GTP
Glucose
Acetyl-CoA
phosphorylation

Oxaloacetate Citrate
NADH+H +
Oxidative

NAD+
28 ATP

NADH+H +
Malate Isocitrate
NAD+
NADH+H +
CO2
Fumarate α-Ketoglutarate
FADH2 NAD+ Amino
FAD CO2
Succinate
Succinyl-CoA acids
GTP, GDP + Pi
Co-A
 Pyruvate

NAD+
PDC
NADH+H+
CO2

Acetyl-CoA

Oxaloacetate Citrate

Malate Isocitrate
1 CO2
Fumarate α-Ketoglutarate
CO2
Succinate
Succinyl-CoA
 1- Isocitrate dehydrogenase
• Isocitrate is oxidatively decarboxylated to α-
ketoglutarate by isocitrate dehydrogenase ( IDH)
• IDH catalyzes the rate-limiting step, as well as the
first NADH-yielding reaction of the TCA cycle
 Pyruvate

NAD+
PDC
NADH+H+
CO2

Acetyl-CoA

Oxaloacetate Citrate

Malate Isocitrate
CO2
Fumarate α-Ketoglutarate
2
CO2
Succinate
Succinyl-CoA
2- α-Ketoglutarate dehydrogenase
The Lovely Co-enzymes For
Nerds
1. Thiamine pyrophosphate
(B1)
2. Lipoic acid
3. CoA (B5, pantothenic acid)
4. FAD (B2, riboflavin)
5. NAD+ (B3, niacin)
 Pyruvate

NAD+
PDC
NADH+H+
CO2

Acetyl-CoA

Oxaloacetate Citrate

Malate Isocitrate
CO2
Fumarate α-Ketoglutarate

Succinate 3 CO2
Succinyl-CoA
 3- Succinyl-CoA synthetase

• conversion of succinyl-CoA to succinate

• Produce GTP
• This reaction is an example of a substrate level
phosphorylation
 Pyruvate

NAD+
PDC
NADH+H+
CO2

Acetyl-CoA

Oxaloacetate Citrate

Malate Isocitrate
CO2
Fumarate α-Ketoglutarate
4
CO2
Succinate
Succinyl-CoA
 4- Succinate dehydrogenase
• Catalyse the oxidation of succinate to fumarate
• This reaction occurs in in inner mitochondrial
membrane
• Requires FAD
• FAD (flavin adenine dinucleotide) is derived
from vitamin riboflavin B2
• Riboflavin deficiency causes lips and mouth
inflammations ( glossitis and stomatitis)
 Pyruvate

NAD+
PDC
NADH+H+
CO2

Acetyl-CoA

Oxaloacetate Citrate

Malate Isocitrate
CO2
Fumarate α-Ketoglutarate
CO2
Succinate
Succinyl-CoA
 Vitamin B1 deficiency: Beriberi

• Thiamine (Vitamin B1)

• Required for oxidative decarboxylation reactions
• Examples include pyruvate dehydrogenase and α-
ketoglutarate dehydrogenase
• Beriberi occurs mostly in people who abuse alcohol
• Excess alcohol impairs vitamin B1 absorption and
storage
• Blood levels of pyruvate, lactate and alanine are
elevated after carbohydrate-rich meal
 Vitamin B2 deficiency

 FAD (Flavin Adenine Dinucleotide)

 Needed for Succinate dehydrogenase
 Deficiency cause inflammation of lips and mouth
( glossitis and stomatitis), scaling and fissures at
the corners of the mouth
 Vitamin B3 deficiency : Pellagra

 NAD (Nicotinamide Adenine Dinucleotide)

 Synthesized from Vitamin B3 (Niacin, Nicotinic Acid)
 Deficiencies result in Pellagra (Dermatitis, Diarrhea,
Dementia)
 Required in relatively high amounts compared to other
vitamins
 Can also be synthesized from tryptophan in the liver
 TCA is amphibolic
• TCA cycle is principally designed to completely oxidize
the carbons of acetyl-CoA to CO2 and H2O
• And to generate large amounts of reduced electron
carriers, NADH and FADH2.
• However, numerous important metabolites are
generated via diversion of intermediates of the cycle
into other biosynthetic pathways
• TCA provides carbon skeletons for gluconeogenesis,
fatty acid synthesis, and interconversion of amino
acids
• Therefore, TCA is amphibolic
Interconnected pathways: TCA with anabolic metabolism

Anaplerotic reactions

Anabolic reactions
 Anaplerotic Reactions
• Anaplerotic reactions replenish intermediates of TCA
cycle as they are removed for synthesis of glucose,
fatty acids, amino acids, or other compounds.
• A key anaplerotic reaction is catalyzed by pyruvate
carboxylase
• Pyruvate carboxylase carboxylates pyruvate forming
oxaloacetate (OAA)
 Anaplerotic reactions

Anaplerotic reactions

Anabolic reactions