Biochemistry final exam

Berglind rnadttir

1. Enzymatic steps and regulation of the pyruvate dehydrogenase enzyme complex.

Summary of the energy synthesis in cells:
Glucose is oxidized in a process called glycolysis. All cells of the body are capable of glycolysis. Under
aerobic conditions the glucose (6-Carbons) is converted to 2 molecules of pyruvate (3-Carbons). The
pyruvate molecules are transported into the mitochondria where it is converted to Acetyl-Coenzyme
A (Acetyl CoA, 2 Carbon). Acetyl CoA then enters the TCA cycle (Tricarboxylic/Citric Acid cycle) by
reacting with the 4-carbon compound Oxaloacetate to form the 6-carbon compound Citrate.
In the Citric Acid Cycle, the hydrogen of the substrate is transferred to the coenzymes NAD+ and
FAD+. The reduced coenzymes donate their hydrogen to the respiratory chain of the inner
mitochondrial membrane. The reoxidation of the reduced coenzymes (NADH, FADH2) is highly
exergonic and the energy created by this process is used to synthesise ATP by oxidative
phosphorylation in the Electron Transport Chain (ETC).
TCA cycle and ETC take place in all cells that contain mitochondria.
Glycolysis participates only in carbohydrate metabolism, however Acetyl CoA is also formed in the
degradation of fatty acids and amino acids. The TCA cycle and the ETC cycle are therefore the final
common pathway for the oxidation of all major nutrients.
Pyruvate dehydrogenase complex
Pyruvate deydrogenase complex is responsible for converting Pyruvate, end product of Glycolysis, to
Acetyl CoA. It is strictly speaking not part of TCA cycle but provides Acetyl Coa which is the substrate
for the cycle. This is an irreversible reaction!
The pyruvate that is formed in glycolysis is formed in the cytosol and needs to be transported into
the mitochondria. It can pass through the outer membrane easily but needs carrier for getting
through the inner membrane.
Pyruvate Dehydrogenase complex is a aggreagate of three enzymes.
1. Pyruvate dehydrogenase (PDH or E1)
2. Dihydrolipoyl Transacetylase (E2) = core enzyme
3. Dihydrolipoyl Dehydrogenase (E3)
They are physicaly linked together which ensures proper sequence without release of intermediates.
The complex is also bound to 2 regulatory enzymes, PDH kinase and PDH phosphatase.
Coenzymes
PDH complex contains 5 coenzymes that act as carriers or oxidants for the intermediates for the
reactions.
E1 requires Thiamin Pyrophosphate(TPP)
E2 requires Lipoic Acid and CoA
E3 requires FAD and NAD+
Way to Remember, Tender Loving Care For Nancy
Enzymatic Steps - 5 steps
1.step-Done By PDH
Oxidative Caboxylation.
Pyruvate reacts with TPP bound to the PDH enzyme and undergoes decarboxylation.
2. step Done by PDH
oxidation
PDH moves 2 electrons and acetyl group from TPP to the core enzyme (E2)

Biochemistry final exam

Berglind rnadttir

3.step Done by E2
Transesterification
SH group of CoA (coenzyme) replaces SH group of E2 to yield Acetyl CoA.
4.Step Done By E3
Oxidation
Enzymes moves 2 Hydrogens atoms from E2 to FAD prosthetic group of E3 forming FADH2
5.step Done by E3
Reduction
The reduced FADH2 group transfers hydrogen to NAD forming NADH.

Role of lipoic acid

Lipoic acid residues, linked to the lysine side chains of dihydrolipoyl transacetylase (E2) participates in
the hydrogen and acyl group transport between subunits and they are involved in acyl group transfer
to CoA.
Regulation
Modification of regulatory enzymes (kinase and phosphatase) activate/inactivate the E1.
Pyruvate dehydrogenase (E1) is inactivated by phosphorylation of 1 single Serine side chain by
cAMP independent PDH Kinase. E1 is reactivated by phosphorylation, which is done by PDH
phosphatase.
The protein kinase is allosterically activated by the same factors that inhibit the pyruvate
dehydrogenase. Pyruvate dehydrogenase is inhibited by NADH and Acetyl CoA, but activated by
AMP.
Thus the PDH kinase is activated by increased NADH/NAD ratio, increased AcetylCoA/CoA ratio and
also ATP. Thos signal high energy state and indicate that there is no need for the cycle to proceed.
The PDH complex is therefore turned off and the TCA cycle activity is reduced.
The PDH kinase in inhibited by Pyruvate. Elevated levels of Pyruvate therefore activate the PDH
complex and turn the TCA cycle back on (or increase it).
2

Biochemistry final exam

Berglind rnadttir

E1 is active dephosphorylated, but inactive when it is phosphorylated.

Calcium (Ca2+) is a strong activator of PDH phosphatase which is important in muscle since contration
increases Ca2+ which then stimulates the PDH complex and increase energy production.

PDH deficiency.
Deficiency of E1 is rare but most common cause of lactic acidosis. The deficiency causes inability to
convert Pyruvate to Acetyl CoA so it gets turned into Lactic Acid via Lactate dehydrogenase. This
causes problems especially for the brain which relies on TCA cycle for energy and is sensitive for
Acidosis. Symptoms include nerve degeneration (since they are deprived of energy).
It is X-linked and affects both male and female.