Regulation of glycolysis

 Why regulation?
 To maintain a constant set of conditions within cells.
 To study related diseases caused by enzymes deficiency and dysfunctions.
 To inhibit and control enzyme functions (such as poisonous substances which can inhibit some
enzymes).

 Key enzymes
The reactions, catalyzed by these enzymes, are irreversible. They push the reactions in only one direction
towards the products, as the reactions towards the substrates are catalyzed by antagonistic enzymes and thus
need to be controlled or regulated.

An Example of antagonistic enzymes within the cell: (Protein Kinase and Protein Phosphatase).

 Hexokinase/Glucokinase
Phosphoryl Transfer Enzyme

The first step in the metabolism of any sugar is phosphorylation, why? Phosphorylation prevents leakage of
the sugar, why? Phosphorylated compounds don’t move or diffuse through the cell membrane, why?
Phosphorylated compounds are trapped in the non-polar part of the cell membrane.

Hexokinase Glucokinase

Site Extrahepatic Liver and pancreatic B cells

Substrate/specificity Low specific for glucose High specific for glucose

Affinity High…why? Low…why?

Km Low… why? High… why?

Vmax Low… why? High… why?

Glucose 6 phosphate effect Allosteric Inhibitor No effect… why?

Feeding/Insulin No effect… why? Induction

Fasting/Anti-Insulin No effect… why? Repression

Km is the concentration of substrate which permits the enzyme to achieve half Vmax. Vmax is the rate of
reaction when the enzyme is saturated with the substrate. Km and Vmax are inverse measures of affinity.
  Phosphofructokinase-1
Phosphoryl Transfer Enzyme

Clarification:

 Fructose 2,6 bisphosphate is the most potent allosteric activator to PFK-1 and the
most potent allosteric inhibitor to fructose 1,6 bisphosphatase (key enzyme for
gluconeogenesis).
 It was found that both PFK-2 and fructose 2,6 bisphosphatase are present in one
enzyme called (Bifunctional Enzyme; an enzyme with two antagonistic functions).
 If this enzyme is dephosphorylated, it is active as PFK-1 and inactive as Fructose
2,6 Bisphosphatase.
 If it is phosphorylated, it is active as Fructose 2,6 Bisphosphatase and inactive as
PFK-2.
 If Fructose 2,6 Bisphosphate is dephosphorylated to (by the action of fructose 2,6
bisphosphatase) or not produced by the phosphorylation of Fructose 6 Phosphate
(by the action of PFK-2), this can indirectly activates fructose 1,6 bisphosphatase.
  Pyruvate kinase
Phosphoryl Transfer Enzyme

This enzyme is active when dephosphorylated and inactive when phosphorylated.

 Other regulatory Mechanisms:

 Energy state of the cell:
1. High level of ATP inhibits PFK-1 and PK.
2. High level of AMP activates PFK-1.

2ATP 2Pi 2ADP Adenylate Kinase ATP + AMP

 Effect of FA oxidation:

It is an important mechanism to inhibit glycolysis during fasting in order to spare glucose for brain and
RBCs, as Fatty acid oxidation can increase level of:

1. ATP which inhibits PFK-1 and PK.

2. Citrate which inhibits PFK-1.
3. Acetyl CoA which inhibits PDH and PK and activates Pyruvate Carboxylase (a potent enzyme in
gluconeogenesis).
  Inhibition of glycolysis according to Pasteur effect:

More availability of O2, more ATP is produced which inhibits the key enzyme phosphofructokinase-1 and
thus decreasing glycolysis. More ATP would usually mean More citrate comes out of the mitochondria and
inhibits the key enzyme phosphofructokinase-1.

 Hormonal regulation:

As mentioned earlier, what are the main mechanisms by which glycolysis is increased (feeding/insulin - 3
mechanisms) and decreased (fasting/anti-insulin - 2 mechanisms)?.

 Importance of glycolysis in RBCs:

1. Glycolysis is The only mechanism for energy production.
2. Glycolysis provides NADH for reduction of met hemoglobin by (NADH met Hb reductase).
3. BPG Shunt.

In this shunt 2,3 bisphosphoglycerate is produced by the action of bisphosphoglycerate mutase and
unites with Hb which helps release oxygen. (Read about Luebering-Rapoport Shunt or cycle or wait to
be studied).

4. Hemolytic anemia due to deficiency of PK enzyme (The second common cause of hemolytic anemia
after glucose 6 phosphate DH; A NADPH generator from glucose 6 phosphate ).

 Important intermediates in glycolysis:

 DHAP can give glycerol 3 phosphate which gives glycerol part of TAG (remember: this reaction is a
pyruvate source as well).
 3 phosphoglycerate can give serine.
 Fate of pyruvate

Alanine
The coenzyme needed for ALT is vitamin B6

Cahill/alanine cycle: this cycle is important for ammonia detoxification in muscles and helps recycle
pyruvate between muscle and liver and get more glucose from pyruvate through gluconeogenesis in liver.

Note: Urea cycle is going to be taken in the next term.

 Another pathway, alpha-KG may take:

Lactate
The coenzyme used for LDH is vitamin B3

Lactate is formed for energy production in the following conditions:

1. Anaerobic state (in the absence of oxygen): it is the most common cause of lactate production.
Clinically, higher lactate concentration would mean hypoxia, ischemia, infection, or heart failure.
2. Absence of mitochondria as in RBCs.
3. PDH metabolic disorder.
4. Cancer.

Lactate formation is the most primary pathway for the following tissues:

Think Lactate When you Can't Make Respirations (Testes, Lens, WBCs, Cornea, Medulla of the Adrenal
gland, and RBCs)

o Remember Cori cycle of lactate between muscle and liver (for glucose production through
gluconeogenesis)

Oxaloacetate
The coenzyme for PC is biotin and Mn is used as a cofactor

Oxaloacetate is used as an available reactant for gluconeogenesis and/or TCA cycle.

Regulation of PC:

1. It is activated by acetyl CoA to ensure the presence of oxaloacetate for TCA cycle as It is a primer
reactant for the cycle.
2. It is activated by anti-insulin as it is a key enzyme for gluconeogenesis.
3. It is inhibited by insulin (Repression effect).
 Acetyl CoA
PDH is a complex enzyme that requires 5 coenzymes (NAD, CoASH, TPP, FA0D, and Lipoic Acid)

Acetyl CoA is produced through oxidative decarboxylation of pyruvate with the production of NADH and
CO2. It is the second primer reactant to proceed TCA cycle.

Regulation of PDH:

1. During exercises: NAD, ADP, and Ca ions are increased and thus PDH is activated.
2. Insulin works directly on Protein Phosphatase to dephosphorylate PDH and thus becomes activated.
3. Any substrate for PDH works as an allosteric activator.
4. Any product of PDH works as an allosteric inhibitor.

Metabolic disorders of PDH:

1. Congenital deficiency of PDH.

2. Thiamin (B1) deficiency.
3. Arsenic poisoning: it binds to thiol group of Lipoic acid.

All of them cause Lactic Acidosis (results in fatal brain damage).

Followed with the regulation for TCA cycle