FADH2

FAD
Reactions of Citric Acid Cycle
1. Citrate synthase: Formation of Citryl CoA intermediate.
Reactions of Citric Acid Cycle
Binding of Oxaloacetate to the enzyme results in conformational change
which facilitates the binding of the next substrate, the acetyl Coenzyme
A. There is a further conformational change which leads to formation of
products. This mechanism of reaction is referred as induced fit model.
– At cellular pH, citric acid is actually present as citrate ion.
Despite this, the name of the cycle is citric acid cycle, which
refers to the molecular, rather than the ionic form of the
substance.
– A synthase is an enzyme that makes a new covalent bond
during a reaction without the direct involvement of an ATP
molecule.
2. Aconitase:
• This enzyme catalyses the isomerization reaction by removing and
then adding back the water (H and OH ) to cis-aconitate in at
different positions.
• Isocitrate is consumed rapidly by the next step thus deriving the
reaction in forward direction.
3. Isocitrate dehydrogenase:
• There are two isoforms of this enzyme, one uses NAD+ and
other uses NADP+ as electron acceptor.
BACK
– The key happenings in step 3 (Isocitrate dehydrogenase) are the
following:
1. The hydroxyl group of isocitrate is oxidized to a ketone group.
2. NAD+ is converted to its reduced form, NADH.
3. A carboxyl group from the original oxaloacetate is removed as
Carbon dioxide.
4. a-Ketoglutarate dehydrogenase:
• This is a complex of different enzymatic activities
similar to the pyruvate dehydogenase complex.
• It has the same mechanism of reaction with E1, E2 and
E3 enzyme units. NAD+ is an electron acceptor.
– The key happenings in step 4:
1. A second NAD+ is converted to its reduced form NADH.
2. A second carboxyl group is removed as carbon dioxide.

3. Coenzyme A reacts with the decarboxylation product succinate to

produce succinyl CoA, a compound with a high energy thioester
bond. This is the second involvement of coenzyme A molecule in
the cycle.
( The other CoA involvement is in step 1)
5. Succinyl CoA synthetase:
• Succinyl CoA, like Acetyl CoA has a thioester bond with very
negative free energy of hydrolysis.
• In this reaction, the hydrolysis of the thioester bond leads to
the formation of phosphoester bond with inorganic
phosphate.
Continuation: This phosphate is transferred to
Histidine residue of the enzyme and this high energy,
unstable phosphate is finally transferred to GDP
resulting in the generation of GTP.
– The thioester bond in Succinyl CoA is a strained
bond.
– Its hydrolysis releases energy, which is trapped
by GTP formation.
– The function of the GTP produced is similar to
that of ATP: to store energy in the form of a
high energy phosphate bond.
6. Succinate Dehydrogenase:
• Oxidation of succinate to fumarate.
• This is the only citric acid cycle enzyme that is tightly bound
to the inner mitochondrial membrane. It is an FAD
dependent enzyme.
• Malonate has similar structure to Succinate, and it
competitively inhibits SDH.
BACK
FADH2
FAD
– Fumarate, with its trans double bond, is an essential
metabolic intermediate in both plants and animals.
– Its isomer, with a cis-double bond, is called maleate,
and it is toxic and irritating to tissues.

– Succinate dehydrogenase produces only the trans

isomer of this unsaturated diacid.
7. Fumarase:
• Hydration of Fumarate to malate: It is a highly
stereospecific enzyme.
• Cis-Maleate (the cis form of fumarate) is not
recognized by this enzyme.
8. L-Malate dehydrogenase:
Oxidation of malate to oxaloacetate:
• It is an NAD+ dependent enzyme.
• Reaction is pulled in forward direction by the next
reaction (citrate synthase reaction) as the oxaloacetate is
depleted at a very fast rate.
SUMMARY OF THE CITRIC ACID CYCLE
Acetyl CoA + 3NAD+ + FAD + GDP + Pi + 2H20 ==

2 CO2 + CoA-SH + 3 NADH + 3H+ + FADH2 +

GTP

*products
Important Cycle Features
1. The “fuel” for the cycle is Acetyl-CoA, obtained from
the breakdown of carbohydrates, fats and proteins.
2. Four of the cycle reactions involve oxidation and
reduction. The oxidizing agent is either NAD(3) or
FAD(1). The operation of this cycle depends on the
availability of these oxidizing agents.
3. In redox reactions, NAD+ is the oxidizing agent when a CARBONOXYGEN double bond
is formed. FAD is the oxidizing agent
when a CARBON-CARBON double bond is formed.
4. The three NADH and one FADH2 that are formed during the
cycle carry electrons and H+ to the ETC through ATP is
synthesized.
5. Two carbon atoms enter the cycle as the acetyl unit of acetyl CoA,
and two carbon atoms leave the cycle as two molecules of CO2.
-The carbon atoms that enter and leave are not the same ones. The
carbon atoms that leave during one turn of the cycle are carbon
atoms that entered during the previous term of the cycle.
6. Four B vitamins are necessary for the proper functioning of the
cycle:
– riboflavin (in both FAD and the alpha ketoglutarate dehydrogenase
complex),

– nicotinamide in NAD,
– pantothenic acid in CoA-SH, and
– thiamine in alpha ketoglutarate dehydrogenase complex.
7. One high-energy GTP molecule is
produced by phosphorylation
CAN
I

CITRATE
ISOCITRATE

KEEP on

α-KETOGLUTARATE

SELLING

SUCCINYL COA

SUBSTANCES

SUCCINATE

FOR

FUMARATE

MONEY,
OFFICER?

MALATE
OXALOACETATE
SUE

SYNTHASE

CITRATE SYNTHASE

AND

ACONITASE

ACONITASE

DANNY

DEHYDROGENASE

DANCE

DEHYDROGENASE

SALSA,

SYNTHETASE

DISCO,

DEHYDROGENASE

FOXTROT FUMARASE
,&
DEHYDROGENASE
DAB

ISOCITRATE
DEHYDROGENASE
α-KETOGLUTARATE
DEHYDROGENASE
SUCCINYL COA SYNTHETASE
SUCCINATE DEHYDROGENASE

FUMARASE
MALATE DEHYDROGENASE
Can

CONDENSATION

ISOMERATION

Open the Door

twice?

OXIDATION & DECARBOXYLATION

Peter

PHOSPHORYLATION

Oliver

OXIDATION

Heads

HYDRATION

Outside.

OXIDATION

BACK

OXIDATION & DECARBOXYLATION