Dr.

Baskaran Krishnan
Associate Professor
Anna Medical College
Mauritius
Specific learning objectives
• Describes various glucose transporters.

• Define glycolysis and its significance.

• Trace glycolytic pathways-aerobic and anerobic with

energetics and regulations.

• Describe Rapaport-Leubering cycle and its significance.

• Describe metabolic fat of pyruvate.

• Describe disorders in glycolytic pathway.

Glucose entry into the cells
 Glucose absorbed are carried by blood.

 Glucose transporters (Glut 1-14) are required for

uptake of glucose by cells from blood stream.

 Glucose enters the cells by two mechanisms

through these transporter.
Glucose entry into the cells…
 1. Insulin independent. Insulin is not required for
the uptake and it occurs in RBC, brain, liver, and
sperms (Glut 1,2,3 and 5).
 2. Insulin dependent. Insulin is required for the
uptake of glucose by the cells and occurs in muscle and
adipose tissue (Glut4).
 In type 2 diabetes mellitus, insulin resistance is seen in
muscle and adipose tissue. Due to defect in Glut4.
 Glucose transporters
Transporters Location Characteristics
GluT1 RBC, brain, kidney, Glucose uptake in colon, retina,
most of cells placenta

GluT2 Serosal surface of Glucose uptake in liver; glucose

intestinal cells, liver, beta sensor in beta cells
cells of pancreas

GluT3 Neurons, brain Glucose into brain cells

GluT4 Skeletal, adipose tissue Insulin mediated muscle, adipose

tissue glucose uptake

GluT5 Small intestine, testis, Fructose transporter; poor ability to

sperms, kidney transport glucose

The other two transporters are SGLT1 and SGLT2. SGLT1 is in SI for
absorption of glucose while SGLT2 is in renal tubules involves in
reabsorption of glucose.
Glucose utilization
 At rest, brain utilizes 80% of glucose in the body.

 During exercise, muscle accounts for 80% of body's

glucose utilization.

 Carbohydrate metabolism is centered at glucose.

 First step is phosphorylation of glucose to glucoe-6-

phosphate.
Glucose utilization…
 This is a trapping mechanism which prevents glucose
from coming out of the cells.

 This is catalyzed by glucokinase in liver and

hexokinase in other tissues.

 They are isoenzymes.

Overview of glucose metabolism
Glycolysis
 It is also known as Embden-Meyerhof Pathway.

 Definition.
 Glucose is split into two molecules of pyruvate (under
aerobic conditions) or lactate (under anaerobic conditions)
by a serries of enzymatic reactions.
Glycolysis…
• Significance.
• It is the major source of energy in brain.
• It is the only source of energy in RBC and fetus and
exercising muscles.
• It can occur in both aerobic and anerobic condition.
• This pathway leads to synthesis of fat through pyruvate.
• Most of the reactions are reversible and used for
gluconeogenesis.
Glycolysis…
 Tissue: In all tissues.

 Intracellular site: cytoplasm.

 End product: Pyruvate in aerobic condition while

lactate in anaerobic conditions.
 Lactate is always the end product in RBC as there is no
mitochondrion and in exercising muscle as there is a
lack of oxygen.
Glycolysis…
 Reactions of glycolysis:
 It has a total of 10 reactions with seven reversible and three
irreversible (non equilibrium) reactions.

 reactions 1, 3 and 10 are irreversible and the rest are reversible.

 Reaction 1: Glu G-6-P catalyzed by GH/HK.

 Reaction 3: F-6-P F-1,6- bisphosphate catalyzed by PFK.

 Reaction 10: PEP Pyruvate catalyzed by PK.

Glycolysis…
 Reactions 7 and 10 generate ATP by substrate level
phosphorylation.
 Reaction 7: 1,3-BPG 3-PG catalyzed by PG kinase.

 Reaction 10: PEP Pyruvate catalyzed by PK.

 The pathway can be divided into two distinct phases.

1. Preparatory or energy investment phase.

2. Pay off phase or energy generation phase.

1. Preparatory or energy investment phase:

 Phosphorylation of glucose and generation of two

molecules of glyceraldehyde-3-phosphate.

 Two mole of ATP are used and no energy produced.

2. Pay off phase or energy generation phase:

 Oxidation of glyceraldehyde-3-phosphate to pyruvate or

lactate with the formation of ATP and NADH.
 Anaerobic glycolysis

Pyruvate
NADH NADH
Lactate
Dehydrogenase (LDH)
NAD+ NAD+

Lactate

• Pathway is the same till pyruvate, then it is reduced by LDH.

• Net energy gain during anaerobic glycolysis is only 2 ATP.
• NADH formed in Gly -3-P DH reaction is utilized by LDH to form NAD+ .
• This NAD+ can now be utilized by Gly-3-P DH so that glycolysis will continue.
Regulation of glycolysis
 Insulin favors glycolysis by activating key glycolytic
enzymes –GH, PFK and PK.

 PFK is rate-limiting enzyme in glycolysis.

 PFK is an allosterically regulated by.

 AMP & fructose-6-phosphate are allosteric activators.

 ATP and citrate are allosteric inhibitors.

Regulatory enzymes of glycolysis
Enzyme Activation Inhibition
HK - G-6-P
GK Insulin Glucagon
PFK Insulin, AMP Glucagon, ATP
F-6-P, PFK-2 Citrate, Low pH
F-2,6-BP Cyclic AMP

PK Insulin, F-1,6-BP Glucagon, ATP

Cyclic AMP

PDH CoA , NAD Acetyl CoA, NADH

Energetics…
Energy yield (number of ATP generated) per molecule of
glucose in the glycolytic pathway, under aerobic conditions
(oxygen is available)
Step Enzyme Source No of ATP gained per
glucose mol
1 Hexokinase - Minus 1
3 Phosphofructokinase - Minus 1
5 Glyceraldehyde-3- NADH 3,0 × 2 = 6
phosphate
dehydrogenase
6 l,3-bisphosphoglycerate ATP 1×2=2
kinase
9 Pyruvate kinase ATP 1×2=2
Total = 10 minus 2 = 8; So net is 8 ATP
Energetics…
Energy yield (number of ATP generated) per molecule of
glucose in the glycolytic pathway, under anaerobic conditions
(Oxygen deficiency)

Step Enzyme Source gained per glucose

mol
1 Hexokinase - Minus 1
3 Phosphofructokinase - Minus 1
6 1,3-bisphosphoglycerate ATP 1x2=2
kinase

9 Pyruvate kinase ATP 1x2=2

Total = 4 minus 2 = 2; So net is 2 ATP
Energetics…
 Number of ATP generated per mol of glucose when it is
completely oxidized through glycolysis and TCA cycle.
 No of ATP formed in the aerobic glycolysis = 10
 No of ATP formed in PDH reaction =6
 No ATP generated in TCA cycle = 24
 Total no of ATP formed = 40
 No of ATP mol used in the priming reactions = -2
 Subtract 2 from 40=38
 So, net generation of number of ATP = 38
Glycolysis erythrocytes
 In RBC, about 25% of glu undergoes an altered glycolysis.

 It is known as Rapaport-Leubering cycle.

 Here 1,3-BPG is directly converted to 2,3-BPG by mutase.

 It is then converted to 3-phosphoglycerate by phosphatase.

 No energy is liberated and energy is dissipated as heat.

 Increase in 2,3-BPG shifts the O2 dissociation curve to the right.

Rapaport-Leubering cycle
 It is also known as BPG shunt in RBC.
 As involves in the synthesis of 2,3-BPG.
 No ATP is synthesized.
Significance of 2,3-BPG
• It combines with Hb and reduces the affinity towards O2.
• So, in presence of 2,3-BPG,oxyhemoglobin will unload
oxygen more easily to tissues.
• In hypoxia, the 2,3-BPG level in the RBC increases.
• It helps release of O2 to the tissues even when pO2 is low.
• In high altitude, O2 density is low, as a result 2,3-BPG
concentration increases.
• This favors release of oxygen to tissues in high altitude.
Fate of pyruvate
 Pyruvate is generated in cytoplasm is transported into
mitochondria by a pyruvate transporter.

 Under aerobic conditions, pyruvate is converted to acetyl

CoA which is oxidized in TCA cycle.

 Acetyl CoA is also used in fatty synthesis.

Fate of pyruvate…
 Pyruvate is carboxylated to oxaloacetate and used for
gluconeogenesis.

 Pyruvate is transaminated to alanine.

 Under anerobic condition, pyruvate is reduced to lactate

by lactate dehydrogenase. (LDH).
Fate of pyruvate…
Pyruvate dehydrogenase (LDH)
• It is multienzyme complex consists of:
• Pyruvate decarboxylase / dehydrogenase.
• Dihydrolipoyl transacetylase.
• Dihydro lipoyl dehydrogenase.

• It requires five coenzymes or cofactors.

• Thiamine pyrophosphate (TPP).
• Lipoamide.
• CoA.
• FAD.
• NAD.
Pyruvate dehydrogenase…
Clinical aspects
 RBC continues to use glucose through glycolysis in
blood collected for blood sugar analysis.

 Sodium fluoride (NaF) is used as preservatives in

blood collection for estimation of sugar.

 This inhibits enolase and thus glycolysis in RBS.

 This ensure that blood sugar values are not lowered.

Clinical aspects…
1. Lactic acidosis may be seen in hypoxia, shock,
pulmonary failure, alcohol abuse.

2. Deficiency of glycolytic enzymes. Very rare.

Pyruvate kinase deficiency & hexokinase deficiency.
They lead to hemolytic anemia and jaundice, because
energy depleted RBCs are hemolyszed.
Clinical aspects…
3.PDH requires TPP.
 TPP deficiency in alcoholism causes pyruvate
accumulation in tissues and resultant lactic acidosis.
 PDH deficiency may also lead to lactic acidosis.
Reflection
• 1.Irreversible reaction of glycolysis are catalyzed by.
1.HK,PFK & PK.
2.GK,PK and glyceraldehyde-3-phosphate dehydrogenase.
3.HK, phosphoglycerate kinase and PK.
4.PK, fructose 1,6-bisphosphatae and PFK.
• 2.Glucose transporter in myocyte stimulated by.
1. GLUT1.
2. GLUT2.
3. GLUT3.
4. GLUT4.