Tricarboxylic acid cycle

(TCA cycle)
Lecture 4
Objectives
1. Define the TCA cycle
2. Identify The Pyruvate Dehydrogenase
complex and its regulation
3. State the stages of the TCA
4. Calculate the energy produced by the TCA
cycle
5. Describe the regulation of the TCA cycle
Steps of glucose oxidation:
1 Glucose
Aerobic
Glycolysis
( in Cytoplasm )

O2 absent (Anaerobic)

2 Pyruvic Acid 2 Lactic Acid

O2 present (Aerobic)

TCA cycle
(In mitochondria) Aerobic Phase
O2 present

6 CO2 + H2O
 Remember the fate of Pyruvate ?

Acetyl CoA √

Pyruvate
Carboxylase
Pyruvate Gluconeogenesis
Oxaloacetate

Lactate
Alanine
√
 1st step
Pyruvate does not enter the TCA cycle directly, but must
be oxidised into Acetyl CoA

Pyruvate
dehydrogenase
Pyruvate Acetyl CoA

TCA cycle
TCA , Krebs cycle,Citric acid cycle

1st step

TCA cycle
 The Pyruvate Dehydrogenase (PDH) Complex

• The PDH complex is a multienzyme complex present in the

mitochondrial matrix.
• It is responsible for oxidative decarboxylation of pyruvic acid
and its conversion to acetyl Co A with formation of NADH + H+
 Pyruvate dehydrogenase complex
(PDHC)
Made of 3 enzymes

Pyruvate Dihydrolipoyl Dihydrolipoyl

decarboxylase transacetylase dehydrogenase

And 5 co-enzyme`

TPP LIPOIC ACID coA FAD NAD

• TPP: Thiamine Pyrophosphate

So the reaction will be:

PDH
• Pyruvate + CoA + NAD+ ------> acetyl-CoA + NADH + H+ + CO2
PDH = pyruvate dehydrogenase

• NADH +H+ is then oxidized in mitochondria (ETC), to produce 3

ATP
 Regulation of the activity of pyruvate
dehydrogenase complex.

1. Allosteric regulation:
– The two products of the complex, NADH and acetyl-CoA, inhibit
the enzyme (negative allosteric effectors).
X
Pyruvate + CoA + NAD+ ----PDH--> acetyl-CoA + NADH + H+ + CO2

2. Covalent modification:
• The phosphorylation of PDH inhibits its activity, while
dephosphorylation activates it.
P

PDHC PDHC
Inactive active
• TCA cycle is a cyclical series of eight reactions
that oxidize one molecule of acetyl CoA
completely to:
– Two molecule of CO2
– Generating energy; either
1. directly in the form of ATP
2. OR indirectly in the form of NADH or FADH2

• The cycle is aerobic (oxygen deficiency inhibit it).

• Site:
– In the mitoc hondrial matrix of all cells containing
mitochondria (but not RBCs because it does not
have mitochondria)
TCA , Krebs cycle,Citric acid cycle

1st step

TCA cycle
 Stages of the TCA cycle
• The cycle can be divided into three stages
according to the role of OAA:
– Stage 1: The attachment of oxaloacetate carrier
(reaction 1)
– Stage 2: Breaking of oxaloacetate carrier (reaction
2-5)
– Stage 3: Regeneration of the oxaloacetate
(reaction 6-8)
Stage 1 of TCA cycle (Formation of citrate)
1: Acetyl-CoA needs a carrier to enter the TCA cycle, this carrier is
oxaloacetate

– Acetyl-CoA + oxaloacetate Citrate Synthase Citrate.

Stage 2 of TCA cycle (Breaking of oxaloacetate)

2: Isomerisation of citrate to isocitrate by Aconitase enzyme
3: Oxidative decarboxylation of isocitrate by Isocitrate Dehydrogenase

Isocitrate Isocitrate Dehydrogenase α -ketoglutarate + CO2

NAD+ NADH + H+
In this reaction:
• The first NADH is produced
• The first CO2 is released
4: Oxidative decarboxylation of α-Ketoglutarate by α-Ketoglutarate
Dehydrogenase Complex:

α-Ketoglutarate α-Ketoglutarate Dehydrogenase succinyl-CoA + CO2

NAD+ NADH + H+
• In this reaction:
– The second NADH is produced
– The second CO2 is released

5: Succinyl CoA is converted to succinate by (Succinate Thiokinase enzyme)

Succinyl CoA Succinate Thiokinase Succinate + CoA

GDP GTP

• GTP is formed from GDP by (substrate-level phosphorylation)

• GTP is then converted to ATP
Stage 3 of TCA cycle (Regeneration of OAA):

6: Oxidation of succinate by succinate Dehydrogenase:

Succinate Succinate dehydrogenase fumarate

FAD FADH2

– FADH2 is oxidized in ETC with production of 2 ATP

7: Fumarase (fumarate hydratase)

Fumarate Fumarase Malate

8: Oxidation of malate by Malate Dehydrogenase (MDH)

Malate Malate Dehydrogenase oxaloacetate

NAD+ NADH + H+
– NADH. is oxidized in respiratory chain to produce 3 ATP
 Energy production in Krebs cycle

One acetyl CoA in one turn of TCA produces

1 ATP bY substrate level phosphorylation succinate

(GTP) thiokinase reaction

bY dehydrogenase reaction (Isocitrate

3 NADH dehydrogenase,alpha-ketoglutarate
3X3=9ATP dehydrogenase,malate dehydrogenase)

1 FADH2 bY dehydrogenase reaction (succinate

1X2=2ATP dehydrogenase)

12 ATP per one Acetyl 12x2=24 ATP per two

CoA Acetyl CoA
Energy produced by Complete oxidation of
glucose through Glycolysis and the Krebs Cycle

Glycolysis 8 ATP
Pyruvate
6 ATP
dehydrogenase
on 2 Pyruvate(2 NADH)
=2x3

Krebs cycle 24 ATP

38 ATP
 Function of Krebs cycle
• It is the final common pathway for oxidation of carbohydrate,
lipids and proteins (as they are metabolized to acetyl CoA or one
of the intermediates of the cycle).

• Oxidation of acetyl CoA with production of ATP.

• In cytosol, citrate gives acetyl CoA which is used in :

– Fatty acid synthesis.
– Cholesterol synthesis.
– Activation of acetyl-CoA carboxylase enzyme (activates fatty
acid synthesis)
– Inhibiton of PFK-1 (inhibit glycolysis)
• Source of the intermediates used in biosynthesis
e.g.
1.Oxaloacetic acid is used in gluconeogenesis.
2.A- ketoglutaric is used for synthesis of some
nonessential amino
3. Succinyl CoA is used in heme synthesis
 Intermediates that used in
other pathways

Gluconeogenesis

In cytosol, citrate gives acetyl CoA

used in synthesis of:Fatty acid
,Cholesterol synthesis.

Nonessential
Aminoacids

Heme synthesis
 Regulation of Citric Acid Cycle
1. Rrespiratory control (energy level in the cell)
•ADP and NAD activate the cycle
• ATP, NADH inhibit the cycle

2. Fine control :
Through regulation of specific enzymes:
•citrate synthase
•isocitrate dehydrogenase, this is the rate-limiting enzyme
•a-ketoglutarate dehydrogenase
These enzymes are inhibited by accumulation of their
products, NADH and ATP
They are allosterically activated by; ADP, AMP and calcium.
Regulation of Citric Acid Cycle
 1 glucose
(Aerobic)
(4 ATP) (Aerobic)
O2 present
O2 present (4 ATP)

pyruvate
pyruvate
1 CO2
NADH (3ATP) 1 CO2
NADH (3ATP)

Acetyl CoA Acetyl CoA

Total ATP = 38
TCA TCA
CYCLE CYCLE
Total CO2= 6

2 CO2+ 12 ATP 2 CO2+ 12 ATP