11/8/2017

Fundamental Medical Science I

Cell metabolism

Lecturer : Dr . Bustanur Rosidi

Learning Objective

Cellular respiration
Carbohydrate metabolism
Fatty acid metabolism

Amino acid metabolism

Nucleotide metabolism

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Glucose

NADH ATP
Pyruvate

NADH CO2
Acetyl CoA

CO2

TCA cycle NADH Electron

transport
chain

FADH
Cellular respiration H+
O2
+
H2O +
ADP + P +
H+
ATP

Glycolysis

Glycolysis :
6
glucose to pyruvate
6

Glucose

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Pyruvate dehydrogenase complex:

CoA

Krebs cycle

Per mol glucose 

6 NADH
2 FADH
2 ATP

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Electron transport chain

The electron transportor respiratory chain gets its name from the fact
electrons are transported to meet up with oxygen from respiration at
the end of the chain.

Electron Transport Chain

NADH
50
• Four protein complexes in 2 e
NAD

the inner mitochondrial

FADH2
Free energy (G) relative to O2 (kcal/mol)

2 e Multiprotein
membrane
FAD
40 FMN
I complexes
Fe S II
Fe S

• A lipid soluble coenzyme

Q
III
Cyt b

(Ubiquinone, CoQ) and a 30 Fe S

water soluble protein (cytc)

Cyt c1 IV
Cyt c

shuttle between protein

Cyt a
Cyt a3
20
complexes
• Electrons generally fall in
energy through the chain -
10 2 e
(originally from
NADH or FADH2)
from complexes I and II to
complex IV 0 2 H + 1/2 O2

H2O

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H
H

Protein H 
H
complex Cyt c
of electron
carriers
IV
Q
III
I
ATP
II synth-
FADH2 FAD 2 H + 1/ O
H2 O
2 2 ase
NADH NAD
ADP  P i ATP
(carrying electrons
from food) H

1 Electron transport chain 2 Chemiosmosis

Oxidative phosphorylation

• The Electron Transport System consists of a chain of electron carriers, associated

with the inner mitochondrial membrane, that bind electrons at successively lower
energy levels. The energy released as the electrons are passed from carrier to carrier
moves hydrogen ions (protons) across the membrane, from the mitochondrial
matrix to the intermembrane space, creating a concentration gradient of protons.

Since the protons carry a charge, an electrical potential (voltage) also develops across the membrane, so the gradient is
often called an electrochemical gradient. This electrochemical gradient is a form of stored energy, some of which is
used to phosphorylate ADP to ATP, a process carried out by a complex of proteins called ATP synthase. As protons move
down their concentration gradient, from the intermembrane space back to the matrix, the energy they release is used
by the ATP synthase complex to phosphorylate ADP.

Complex I receives electrons from NADH, whereas complex II receives them from FADH2. Complexes III
and IV are further down the chain, and ultimately transfer the electrons to molecular oxygen to form
water.
In addition to these integral complexes, two smaller carriers play critical roles. Ubiquinone, a low-
molecular-weight compound within the membrane, receives electrons from complexes I and II, and
transfers them to complex III. Cytochromec, a small peripheral protein, receives electrons from complex
III and transfers them to complex IV. Several other cytochromes are included as members of the four
electron-carrier complexes. Cytochromes are a class of small proteins containing heme that are
important in transferring electrons in cellular processes.

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The total yield of ATP from 1 glucose molecule going through glycolysis and the citric acid
cycle is:
* 2 ATP generated in glycolysis directly.
* 2 NADH generated in glycolysis = 6 ATP (by oxidative phosphorylation).
* 2 NADH generated when 2 pyruvate are converted to 2 acetyl CoA = 6 ATP
(by oxidative phosphorylation)
* 2 acetyl CoA enter citric acid cycle to generate

* 6 NADH = 18 ATP (by oxidative phosphorylation)

* 2 FADH2 = 4 ATP (by oxidative phosphorylation)
* 2 GTP = 2ATP

Total number of ATP from 1 molecule of glucose = 2 + 6 + 6 + 18 + 4 + 2 = 38.

Aerobic vs anaerobic respiration

Aerobic = oxygen is present

Anaerobic = no oxygen

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ATP generation from anaerobic glycolysis

During exercise, like running, muscle can perform anaerobic glycolysis
where pyruvate is further convert into lactate.

Glucose
2ATP

NAD+ NAD+

NADH NADH
4ATP

Lactate Pyruvate Pyruvate Lactate

ATP generation from anaerobic glycolysis

During exercise, like running, muscle can perform anaerobic glycolysis
where pyruvate is further convert into lactate.

• During strenuous exercise, oxygen is in short supply.

• The middle carbonyl in pyruvate is reduced to an
alcohol group, and lactate is formed.
• NADH supplies the hydrogen needed in the reaction.
NAD+ is produced.
• NAD+ is funneled back into glycolysis to oxidize more
glucose. No energy is produced

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Lactate Recycling

• How your liver helps you during exercise....

• Recall that vigorous exercise can lead to a buildup of lactate
and NADH, due to oxygen shortage and the need for more
glycolysis
• NADH can be reoxidized during the reduction of pyruvate to
lactate
• Lactate is then returned to the liver, where it can be reoxidized
to pyruvate by liver LDH
• Liver provides glucose to muscle for exercise and then
reprocesses lactate into new glucose

ATP generation from anaerobic glycolysis

Red blood cells do not have

mitochondria.
ATPs in the RBC are supplied via
anaerobic glycolysis.

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Cori cycle
6 ATP
glucose

Gluconeogenesis
glucose

2 lactate glucose

2 ATP Glycolysis
2 lactate
2 lactate

Cori cycle
6 ATP glucose
Gluconeogenesis
glucose
2 lactate glucose
2 ATP Glycolysis
2 lactate
2 lactate
Lactate generated from glucose by anaerobic glycolysis in the skeletal muscle during
exercise is moved to the liver, reoxidized to pyruvate and turned back into glucose by
gluconeogenesis, which is then returned to the muscle or other peripheral tissues

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Fatty acids Glucose

NADH ATP
NADH
Pyruvate

NADH CO2
FADH
Acetyl CoA

CO2

TCA cycle NADH Electron

transport
chain

FADH
Cellular respiration H+
O2
+
H2O +
ADP + P +
H+
ATP

Words to know
 Adenosine Triphosphate
 Metabolism
 Catabolism
 Anabolism
 Coenzymes
 Glycolysis
 Pyruvate
 Lactate
 Acetyl CoA
 TCA Cycle
 Electron Transport Chain
 Oxydative Phosphorylation
 Beta Oxidation
 Deamination&Transamination

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Part I

Cell metabolism - II

Carbohydrate metabolism
Fatty acid metabolism

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Cellular respiration

Glycolysis
TCA cycle
OxydativePhosphorylation
β-Oxydation

Fatty acids Glucose

NADH ATP
NADH
Pyruvate

NADH CO2
FADH
Acetyl CoA

CO2

TCA Electron
NADH
cycle transport
chain

FADH

Cellular respiration H+
O2
+
H2O +
ADP + P +
H+
ATP

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Carbohydrate metabolism

 Gluconeogenesis
 Fructose
 Galactose
 Glycogen
 Pentose phosphate pathway

Cori cycle
6 ATP
glucose

Gluconeogenesis
glucose

2 lactate glucose

2 ATP Glycolysis
2 lactate
2 lactate

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Gluconeogenesis
G
L
Y
C
Going in reverse
O
L
I
S
I
S

Gluconeogenesis
G
L
Y
C
O
From cleavege of L
Triacylglycerol (fatt)
I
S
I
S

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Gluconeogenesis
Synthesis of "new glucose" from common metabolites

Humans consume 160 g of glucose per day

75% of that is in the brain

Body fluids contain only 20 g of glucose

Glycogen stores yield 180-200 g of glucose

So the body must be able to make its own glucose

Gluconeogenesis
Synthesis of "new glucose" from non-carbohydrate sources

Why necessary?
Brain depends on a constant glucose supply. Hypoglycemia
inevitably leads to unconsciousness and damage to the nerve cells
which will die without glucose supply.
Glycogen storage in liver is sufficient for 24 h only, thus body has to
synthesize glucose.

Glugoneogenesis uses pyruvate to make glucose.

Other tissue depending on glucose : RBC, retina cells and medulla of

the kidneys

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Pyruvate carboxylase

Pyruvate carboxylase is a
compartmentalized reaction. Pyruvate is
converted to oxaloacetate in the
mitochondria.
Because oxaloacetate cannot be
transported across the mitochondrial
membrane, it must be reduced to
malate, transported to the cytosol, and
then oxidized back to oxaloacetate before
gluconeogenesis can continue.

 Here : Oxaloacetate needed for TCA cycle is produced !!

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Substrates for Gluconeogenesis

• Pyruvate, lactate, glycerol,

amino acids and all TCA
intermediates can be utilized

Regulation: when ATP or

acetyl-CoA are high, pyruvate
enters gluconeogenesis

Glucogenic Aminoacids

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Last Step in Gluconeogenesis

Conversion of Glucose-6-P to Glucose
• Enzyme: Glucose-6-Phosphatase
• Presence of G-6-Pase in ER of liver and kidney cells makes gluconeogenesis
possible (embedded in the ER membrane, with catalytic site exposed to ER lumen)
• Muscle and brain do not do gluconeogenesis
• G-6-P is hydrolyzed as it passes into the ER
• ER vesicles filled with glucose diffuse to the plasma membrane,
fuse with it and open, releasing glucose into the bloodstream.

How Is Gluconeogenesis Regulated?

• Reciprocal control with glycolysis

• When glycolysis is turned on, gluconeogenesis should
be turned off
• When energy status of cell is high, glycolysis should be
off and pyruvate, etc., should be used for synthesis and
storage of glucose
• When energy status is low, glucose should be rapidly
degraded to provide energy

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Carbohydrate metabolism

Fructose
Galactose
Glycogen
Pentose phosphate pathway

Are Substrates Other Than Glucose Used in Glycolysis?

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Sugar metabolism

small medium large

SSucrose MMaltose LLactose

Glucose Glucose Glucose
+ + +
FFructose GGlucose Gal
Galactose

Sugar metabolism Glycogen

Glucose Glucose-6-P Glucose-1-P UDP-glucose

UDP-galactose

Fructose 6-P
UDP-glucose

Fructose 1,6-biP Galactose 1-P

DiHidroxyAcetone P Galactose
Glyceraldehyde 3-P
Aldolase B
Glyceraldehyde Fructose 1-P
Fructokinase Fructose

Pyruvate

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Sugar metabolism

small

SSucrose
Glucose
+
FFructose

Sugar metabolism: Fructose

Glucose
Fructose
Polyol pathway

Dietary sucrose

Dietary fructose

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Sugar metabolism: Fructose Fructokinase

Liver
Small intestinal mucose
Peroximal epithelium of renal tubule
Glucose Glucose-6-P
Not found in liver
Aldosereductase
Fructose 6-P
Sorbitol

Fructose 1,6-biP Sorbitol

dehydrogenase
Not found in liver

Glyceraldehyde 3-P DiHidroxyAcetone P

Aldolase B
Glyceraldehyde Fructose 1-P
Fructokinase Fructose

Pyruvate

Polyol pathway Aldose reductase: low affinity

to glucose at normal
condition NADPH
Hyperglycemia: excessive amount of glucose in blood plasma
NADP
Glucose Glucose-6-P
Aldose reductase

The lens, peripheral nerves, and

renal glomeruli are insulin insensitive Sorbitol

Blood glucose NAD

Sorbitol
dehydrogenase

Insulin
NADH
Polyol pathway ON

Fructose
Glycogen synthase
Glucose Glycogen Glucose

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Polyol pathway
NADPH
Hyperglycemia: excessive amount of glucose in blood plasma
NADP
Glucose Glucose-6-P
Aldose reductase

Sorbitol
Sorbitol
NAD

Sorbitol
dehydrogenase
Glycate nitrogen
on protein NADH

Fructose
Collagen formation
(cataract)

Sorbitol is formed in many tissues when blood glucose levels are

increased. This occurs through the "polyol pathway", a reaction
sequence found in most tissues. Glucose is first reduced to
sorbitol in the aldolse reductase reaction. It is then oxidized to
fructose by sorbitol dehydrogenase.

The high blood glucose concentrations found in diabetic patients

activate the aldolase reductase enzyme, leading to sorbitol
formation in several organs, notably the kidneys, nerves and the
lens of the eye. Sorbitol accumulates in these tissues because they
lack a membrane transport system for this sugar. They can reduce
glucose, but cannot utilize or get rid of the resulting sorbitol.

Cataract formation in the lens is believed to follow sorbitol

accumulation. Renal damage and neuropathy seen in diabetic
patients is thought to be at least partially triggered by sorbitol
accumulation in these tissues.

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Fructose intolerance (aldolase B deficiency)

Glucose Glucose-6-P

Fructose 6-P

Fructose 1,6-biP

Glyceraldehyde 3-P DiHidroxyAcetone P

Aldolase B
Glyceraldehyde Fructose 1-P
Fructokinase Fructose

Pyruvate
Liver phosphorylase
(glycogen phosphorylase)

Fructose intolerance (aldolase B deficiency)

Glucose Glucose-6-P

Fructose 6-P

Fructose 1,6-biP

Glyceraldehyde 3-P DiHidroxyAcetone P

AldolaseB
Glyceraldehyde Fructose 1-P
Fructokinase Fructose

Pyruvate
Liver phosphorylase
(glycogen phosphorylase)

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Fructose intolerance (aldolase B deficiency)

Fructose
Glucose1-P
mobilization by
glycogen
degradation
Liver phosphorylase
(glycogen phosphorylase)

Glycogen degradation

Glucose availability

Blood glucose
 accumulation of F1P, high levels of F1P traps
phosphate in an unusable form
 Glycogenolysis stops

Aldolase B plays a key role in carbohydrate metabolism as it

catalyzes one of the major steps of the glycolytic-gluconeogenic
pathway. Though it does catalyze the breakdown of glucose, it plays
a particularly important role in fructose metabolism, which occurs
mostly in the liver, renal cortex, and small intestinal mucosa.

When fructose is absorbed, it is phosphorylated by fructokinase to

form fructose 1-phosphate (F1P). Aldolase B then catalyzes F1P
breakdown into glyceraldehyde and DHAP. After glyceraldehyde is
phosphorylated by triosekinase to form G3P, both products can be
used in the glycolytic-gluconeogenic pathway, that is, they can be
modified to become either glucose or pyruvate.

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Genetic mutations leading to defects in aldolase B result in a

condition called hereditary fructose intolerance (HFI). Due to the
lack of functional aldolase B, organisms with HFI cannot properly
process F1P, which leads to an accumulation of F1P in bodily
tissues. In addition to being toxic to cellular tissues, high levels of
F1P traps phosphate in an unusable form that does not return to
the general phosphate pool, resulting in depletion of both
phosphate and ATP stores. The lack of readily available phosphate
causes the cessation of glycogenolysis in the liver, which results in
hypoglycemia. This accumulation also inhibits gluconeogenesis,
further reducing the amount of readily available glucose. The loss
ATP leads to a multitude of problems including inhibition of protein
synthesis and hepatic and renal dysfunction. Patient prognosis,
however, is good in cases of hereditary fructose intolerance. By
avoiding foods containing fructose, sucrose, and sorbitol, patients
can live symptom-free lives.

Carbohydrate metabolism

Fructose
Galactose
Glycogen
Pentose phosphate pathway

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Lactose = galactose + glucose

Lactose intolerance

Intestinal lumen
Lactase deficient cell
Bacterial fermentation
Lactic acid
Osmotic effect
Fluid load
Peristaltic
Watery diarrhea

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Symptoms occur because the unabsorbed lactose passes through the small intestine and into
the colon. In the colon, one type of normal bacterium contains lactase and is able to split the
lactose and use the resulting glucose and galactose for its own purposes. Unfortunately, when
they use the glucose and galactose, these bacteria also release hydrogen gas. Some of the gas is
absorbed from the colon and into the body and is then expelled by the lungs in the breath. Most
of the hydrogen, however, is used up in the colon by other bacteria. A small proportion of the
hydrogen gas is expelled and is responsible for the increased flatulence (passing gas). Some
people have an additional type of bacterium in their colons that changes the hydrogen gas into
methane gas, and these people will excrete only methane or both hydrogen and methane gas in
their breath and flatus.
Not all of the lactose that reaches the colon is split and used by colonic bacteria. The unsplit
lactose in the colon draws water into the colon (by osmosis). This leads to loose, diarrheal
stools.
The severity of the symptoms of lactose intolerance varies greatly from person to person. One
reason for this variability is that people have different amounts of lactose in their diet; the more
lactose in the diet, the more likely and severe the symptoms. Another reason for the variability
is that people have differing severities of lactase deficiency, that is, they may have mild,
moderate, or severe reduction in the amounts of lactase in their intestines. Thus, small amounts
of lactose will cause major symptoms in severely lactase deficient people but only mild or no
symptoms in mildly lactase deficient people. Finally, people may have different responses to the
same amount of lactose reaching the colon. Whereas some may have mild or no symptoms,
others may have moderate symptoms. The reason for this is not clear but may relate to
differences in their intestinal bacteria.

Carbohydrate digestion

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Sugar metabolism Glycogen

Glucose Glucose-6-P Glucose-1-P UDP-glucose

UDP-galactose

Fructose 6-P
UDP-glucose

Fructose 1,6-biP Galactose 1-P

DiHidroxyAcetone P Galactose
Glyceraldehyde 3-P

AldolaseB
Glyceraldehyde Fructose 1-P
Fructokinase Fructose
Pyruvate

Galactose

Glucose Glucose-6-P Glucose-1-P

UDP-galactose

Fructose 6-P Galactose 1-P

uridylyltransferase
UDP-glucose

Fructose 1,6-biP Galactose 1-P

Galactokinase

Galactose
Glyceraldehyde 3-P DiHidroxyAcetone P

Pyruvate

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Classical galactosemia

Glucose Glucose-6-P Glucose-1-P

UDP-galactose

Fructose 6-P Galactose 1-P

uridylyltransferase
UDP-glucose

Fructose 1,6-biP Galactose 1-P

Galactokinase

Galactose
Glyceraldehyde 3-P DiHidroxyAcetone P

Pyruvate

Non-classical galactosemia

Glucose Glucose-6-P Glucose-1-P

UDP-galactose

Fructose 6-P Galactose 1-P

uridylyltransferase
UDP-glucose

Fructose 1,6-biP Galactose 1-P

Galactokinase

Galactose
Glyceraldehyde 3-P DiHidroxyAcetone P

Pyruvate

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Galactosemia

Type II

Type III
Type I

Galactose metabolism

Type II

Type I
Type III

Type I : GALT
Type II : GALK
Type III : GALE

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Alternate Galactose metabolism

Classical galactosemia
If infants with classic galactosemia are not treated promptly with a low-galactose
diet, life-threatening complications appear within a few days after birth. Affected
infants typically develop feeding difficulties, a lack of energy (lethargy), a failure to
gain weight and grow as expected (failure to thrive), yellowing of the skin and whites
of the eyes (jaundice), liver damage, and bleeding. Other serious complications of
this condition can include overwhelming bacterial infections (sepsis) and shock.
Affected children are also at increased risk of delayed development, clouding of the
lens of the eye (cataract), speech difficulties, and intellectual disability. Females with
classic galactosemia may experience reproductive problems caused by ovarian
failure.

Galactosemia type II causes fewer medical problems than the classic type. Affected
infants develop cataracts, but otherwise experience few long-term complications.
The signs and symptoms of galactosemia type III vary from mild to severe and can
include cataracts, delayed growth and development, intellectual disability, liver
disease, and kidney problems.

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Carbohydrate metabolism

Fructose
Galactose
Glycogen
Pentose phosphate pathway

Metabolic homeostasis

Fuel
Blood stores
fuel

Dietary fuels: Blood

-Carbohydrate fuel
-Fat
-Protein

Fuel utilization

Blood ATP
fuel

Cell function

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Glycogen in liver: maintaining blood glucose

Glycogen

Glucose-1-P
Glucose
Glucose-6-P

Blood glucose

Glycogen in muscle: ATP source

Glycogen

Glucose-1-P

Glucose-6-P

Lactate
ATP
CO2

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Glycogen = fuel reserve

The liver and muscles are the major storage place for this energy
reserve

The liver glycogen  sufficient only for an overnight fast

During long period of fasting  use of the fat reserves

The brain cannot use fat directly as a fuel, thus it is dependent upon a
steady supply of glucose via the blood

If the brain is denied glucose, it ceases to function properly

Low plasma glucose level can cause dizziness, faintness, lethargy

Glycogen

Glycogen is the storage form of glucose

6

found in most types of cells.

1

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Glycogen

It is composed of glucosyl units

6

linked by -1,4 glycosidic bonds, with -1,6 branches

occurring roughly every 8 to 10 glucosyl units. 1

Glycogen synthase catalyze the synthesis of glycogen

Blood glucose

Glycogen = fuel reserve

Insulin

Glycogen synthase
Glucose Glycogen

• Glycogen is made from many glucose molecules joined together to form a compact
spherical structure.
 If cell is supplied with surplus glucose, the mitochondria is in high energy state, the
capacity for metabolic flux through TCA Cyle is overwhelmed and certain
metabolites e.g. citrate, ATP accumulate.
 In liver and muscle, some of the excess glucose is channeled along the metabolic
pathway to Glycogen

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Glycogen synthase is activated by insulin

IRS: Insulin Receptor Substrate

Glycogen synthase is activated by insulin

PI-3K: phosphoinositide 3-kinase PKB: Protein Kinase B

PIP2: phosphatidylinositol 4,5-bisphosphate IRS: Insulin Receptor Substrate
PIP3: phosphatidylinositol 3,4,5-trisphosphate GSK: Glycogen Synthase Kinase

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Sugar metabolism Glycogen

Glucose Glucose-6-P Glucose-1-P UDP-glucose

UDP-galactose

Fructose 6-P
UDP-glucose

Fructose 1,6-biP Galactose 1-P

DiHidroxyAcetone P Galactose
Glyceraldehyde 3-P

AldolaseB
Glyceraldehyde Fructose 1-P
Fructokinase Fructose

Pyruvate

Glycogen metabolism Glycogen

Glycogen synthase

Glucose Glucose-6-P Glucose-1-P UDP-glucose

Fructose 6-P

Fructose 1,6-biP

Glyceraldehyde 3-P DiHidroxyAcetone P

Pyruvate

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Glycogen synthesis

Glycogen

Glycogen synthase

Glucose Glucose-6-P Glucose-1-P UDP-glucose

Glycogen synthesis

Glycogen

Glycogen synthase

Glucose Glucose-6-P Glucose-1-P UDP-glucose

UTP PPi

Uridine diphosphate glucose (UDP-glucose) is the immediate precursor for

glycogen synthesis.
Nucleotide diphosphate sugars are precursors also for synthesis of other
complex carbohydrates, including oligosaccharide chains of glycoproteins, etc.

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Glycogen synthesis

• Glycogenin (a protein!) forms the core of a

glycogen particle
• First glucose is linked to a tyrosine -OH
• Glycogen synthase transfers glucosyl units
from UDP-glucose to C-4 hydroxyl at a
nonreducing end of a glycogen strand.

Glycogen degradation

The enzyme starts at the end of a chain

and successively cleaves glucosyl residues

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Glycogen degradation

2 1 Removes 3 glucose residues, and

adds it to the end of a longer chain
by an -1,4 bond.

2 Hydrolyze the -1,6-bond

Hormonal Regulation of Glycogen Synthesis and Degradation

Glycogen breakdown Glycogen synthesis

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Hormonal Regulation of Glycogen Synthesis and Degradation

Glycogen breakdown Glycogen synthesis

Hormonal Regulation I
• of Glycogen Synthesis and Degradation
• Insulin is secreted from the pancreas (to liver) in response
to an increase in blood glucose
• Insulin stimulates glycogen synthesis and inhibits glycogen
breakdown

Hormonal Regulation II
• Glucagon and epinephrine
• Glucagon and epinephrine stimulate glycogen breakdown -
opposite effect of insulin!
• Glucagon is also secreted by pancreas
• Glucagon acts in liver and adipose tissue only
• Epinephrine (adrenaline) is released from adrenal glands
• Epinephrine acts on liver and muscles

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Epinephrine and Glucagon

• The difference....
• Both are glycogenolytic but for different reasons!
• Epinephrine is the fight or flight hormone
– rapidly mobilizes large amounts of energy
• Glucagon is for long-term maintenance of steady-
state levels of glucose in the blood
– activates glycogen breakdown
– activates liver gluconeogenesis

Carbohydrate metabolism

Fructose
Galactose
Glycogen
Pentose phosphate pathway

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Fatty acid synthesis < NADPH NADP

CO2
Ribulose 5-P Glucose-6-P
Oxidative Glucose

Xyulose 5-P
Fructose 6-P

Non-oxidative
Ribose 5-P
Fructose 1,6-biP

Nucleotide synthesis

Glyceraldehyde 3-P DiHidroxyAcetone P

Pyruvate

Pentose phosphate pathway

Glucose Glycolysis
ATP
Glucose 6-Phosphate or

Pentose Phosphate Pathway

In a well-fed state, Glucose is phosphorylated to Glucose-6 phosphate
and it enters the pentose phosphate pathway.
• Provides NADPH for biosynthesis
• Produces ribose-5-P Nucleotide biosynthesis
• Two irreversible oxidative processes followed by five non-oxidative steps
• NADPH is used in cytosol for fatty acid synthesis

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Clinical Aspect
Glucose-6-P-Dehydrogenase Deficiency

Clinical Aspect
Glucose-6-phosphate
NADP
Reduced Glucose-6-P-dehydrogenase
NADPH
Glutathione
6-phosphoglucono-δ-lactone

6-phosphogluconate
NADP Phosphogluconate DH

Reduced
NADPH CO2
Glutathione
Ribulose-5-phosphate

Ribose-5-phosphate

The glucose 6-phosphate DH (G6PD)

reaction is the rate limiting step
and is essentially irreversible

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Glucose-6-phosphate
NADP
Reduced Glucose-6-P-dehydrogenase
NADPH
Glutathione
6-phosphoglucono-δ-lactone

6-phosphogluconate

Ribulose-5-phosphate

Ribose-5-phosphate

Glucose-6-phosphate
NADP
Reduced Glucose-6-P-dehydrogenase
NADPH
Glutathione
6-phosphoglucono-δ-lactone

6-phosphogluconate
NADP
Reduced
NADPH CO2
Glutathione
Ribulose-5-phosphate

Ribose-5-phosphate

The glucose 6-phosphate DH (G6PD)

reaction is the rate limiting step
and is essentially irreversible
Phosphogluconate DH

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NADPH reduces Glutathione

• Reduced Glutathiones function as antioxidant (scavenger for

dangerous oxidative metabolites )
• Reduced glutathione (GSH) maintains the normal reduced state of
the cell.
• The enzyme glutathione reductase uses NADPH as a cofactor to
reduce GSSG back to two moles of GSH. Thus, the pentose pathway
is linked to the supply of adequate amounts of GSH.

NADPH reduces Glutathione

2 NADPH 2NADP+

Reduced glutathione
(GSH)
It is a g-Glu-Cys-Gly

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Glutathione and Erythrocytes

• Mature RBCs have no mitochondria and are totally dependent on NADPH from
the PPP to regenerate GSH from GSSG via glutathione reductase.
• In RBC 10% of glucose consumption goes to pentose pathway.
• Reduced GSH maintains cysteine residues in hemoglobin and other proteins in a
reduced state.
• GSH is essential for normal RBC structure and keeping hemoglobin in Fe++ state.
• Reduced glutathione also detoxifies peroxides (and free radicals).
2GSH + ROOH  GSSG + H2O + ROH
• Cells with low levels of GSH are susceptible hemolysis.
• Individuals with reduced GSH are subject to hemolysis.
• This is often clinically seen as black urine under certain conditions.
• The ingestion of oxidative agents that generate peroxides or reactive oxygen
species (ROS) (e.g Antimalaria – pamaquine, purine glycoside from fava beans).
• Individuals with G6PD deficiency can not produce sufficient GSH to cope with
the ROS. Proteins become cross linked leading to Heinz body formation and cell
lysis.

G6PDH deficiency is an X-linked recessive hereditary disease, and the most common
human enzyme defect, being present in more than 400 million people worldwide

G6PDH is responsible for maintaining adequate levels of NADPH inside the cell. NADPH is a
required cofactor in many biosynthetic reactions and also provides electrons to reduce
oxidized glutathione (GSSG) to reduced glutathione (GSH) catalyzed by glutathione
reductase.

Reduced glutathione acts as a scavenger for dangerous oxidative metabolites in the cell;
in particular, it converts harmful hydrogen peroxide to water with the help of the enzyme
glutathione peroxidase

G6PDH deficiency leads to reduced level of NADPH, and consequently the ability of cell to
cope with oxidative stress. This has a profound effect on the stability of red blood cells
since they are especially sensitive to oxidative stresses

The accumulation of hydrogen peroxide affects many intracellular processes and results in
hemolysis. These include the cross-linking of membrane proteins; hemoglobin
denaturation, which in turn affects the physical properties of the erythrocyte; and lipid
peroxidation, which may affect the cell membrane to cause direct hemolysis.

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G6PD Deficiency can lead to

jaundice

Neonatal screening for G6PD

Deficiency in Hong Kong

www.fhs.gov.hk

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Fatty acid metabolism

b-oxidation
Fatty acid synthesis

Fatty acids Glucose

NADH ATP
NADH
Pyruvate

NADH CO2
FADH
Acetyl CoA

CO2

TCA cycle NADH Electron

transport
chain

FADH
Cellular respiration H+
O2
+
H2O +
ADP + P +
H+
ATP

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The carbon in fatty acids (mostly CH2) is reduced (so its oxidation yields the
most energy possible)

Fatty acids are not hydrated (as mono- and polysaccharides are), so they can
pack more closely in storage tissues

Triacylglycerols are a major form

of stored energy in animals

The energy available in stores

C O C O C O

of fat in average person far

exceeds the energy available
from protein, glycogen and
glucose

Fat is stored mainly as

triacylglycerols in specialized
cells called adipocytes or
adipose cells

Triacylglycerol / Triglyceride

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Liberation of fatty acids from

triacylglycerols in adipose tissue is
hormone-dependent (e.g.
Glucagon, Adrenaline)

COO Fatty acid: ATP generation

Free fatty acids CoA

CoA ATP
FAD
AMP
FADH
NAD
Acyl CoA b-oxidation
NADH H+ H+
Electron
O2 transport
Acetyl CoA chain
H2O

TCA
ADP ATP synthase
+ Pi
H+
ATP

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b-oxidation O
b a
CH3 CH2 CH2 CH2 C SCoA
Fatty acyl CoA FAD
FADH
NAD
NADH
O O
a
CH3 CH2 C CH2 C SCoA
b
CoASH
Acetyl CoA
Fatty acyl CoA
O O
b a a
CH3 CH2 CH2 C SCoA CH3 C SCoA

b -Oxidation of Fatty Acids

Products: acetyl-CoA and a fatty acid chain two carbons shorter

The b-oxidation of saturated
fatty acids involves a cycle of
four enzyme-catalyzed
reactions. Each cycle produces
single molecules of FADH2,
NADH, and acetyl-CoA and yields
a fatty acid shortened by two
carbons
Repetition of the cycle yields
a succession of acetate units
Thus, palmitic acid (C16) yields
eight acetyl-CoAs
These eight Acetyl-CoAs enter
then the TCA cycle and
subsequent Oxydative
Phosphorylation

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The total ATP yield from oxidation of palmitate (C16)

* 8 acetyl CoA enter citric acid cycle and give:
24 NADH = 72 ATP (by oxidative phosphorylation)
8 FADH2 = 16 ATP (by oxidative phosphorylation)
8 GTP = 8 ATP
* 7 NADH generated by beta oxidation itself = 21 ATP (by oxidative phosphorylation)
* 7 FADH2 generated by beta oxidation itself = 14 ATP (by oxidative phosphorylation)

Total number of ATP from 1 molecule of palmitate = 72 + 16 + 8 + 21 + 14 = 131 (-2 ATP

from the free fatty acid activation to AcylCoA).

The total yield of ATP from 1 glucose molecule going through glycolysis and the citric acid
cycle is:
* 2 ATP generated in glycolysis directly.
* 2 NADH generated in glycolysis = 6 ATP ((by oxidative phosphorylation).
* 2 NADH generated when 2 pyruvate are converted to 2 acetyl CoA = 6 ATP
(by oxidative phosphorylation)
* 2 acetyl CoA enter citric acid cycle to generate

* 6 NADH = 18 ATP (by oxidative phosphorylation)

* 2 FADH2 = 4 ATP (by oxidative phosphorylation)
* 2 GTP = 2ATP
Total number of ATP from 1 molecule of glucose = 2 + 6 + 6 + 18 + 4 + 2 = 38.

Fatty acid metabolism

b-oxidation
Fatty acid synthesis

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Differences between fatty acid biosynthesis and breakdown

• b-oxidation is in mitochondria
• Fatty acid biosynthesis is in
cytosol
• Biosynthesis uses NADPH/NADP+;
breakdown uses NADH/NAD+
• Biosynthesis starts with Acetyl-
CoA and Malonyl-CoA
• Acetyl-CoA is transported to
cytoplasm as citrate
cytosol

mitoch

Both oxidation and synthesis of fats utilize an activated

two carbon intermediate, acetyl-CoA.
However, the acetyl-CoA in fat synthesis exists temporarily
bound to the enzyme complex as malonyl-CoA

The synthesis of malonyl-CoA is the first committed step

of fatty acid synthesis and the enzyme that catalyzes
this reaction, acetyl-CoA carboxylase (ACC),
co-factor.

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Oxaloacetate

Fatty Acid Synthase

ACC-Phosphate is inactive.
Insulin activates a phosphatase
which activates ACC

Acetyl CoA Malonyl-CoA

Oxaloacetate

NADPH
Fatty Acid
Synthase NADP

Malonyl-CoA
Oxaloacetate Fatty Acid
Synthase
Acetyl
= ACC

Fatty Acid Synthase

Palmitate (C16)

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Regulation of fatty acid synthesis

Acetyl CoACarboxylase (ACC) plays a key role in fatty acid synthesis

Glucagon stimulation result in phosphorylation and inhibition of ACC
Insulin leads to activation of phosphatases, thereby leading
to dephosphorylation of ACC that results in increased ACC activity

Differences between fatty acid biosynthesis and breakdown

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Synthesis of fatty acid and triacylglycerol

Triacylglycerol

VLDL: very low density lipoprotein

Synthesis of fatty acid and triacylglycerol

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lipoprotein lipase (LPL)

low-density lipoproteins (LDL)

high-density lipoproteins (HDL)

Dietary cholesterol and triglycerides are packaged into chylomicrons in the

intestine, before passing into the bloodstream via lymphatics. Chylomicrons
are broken down by lipoprotein lipase (LPL) in the capillaries of muscle and
adipose tissue to fatty acids, which then enter the cells. The chylomicron
remnants, which have lost much of their triglyceride content, are taken up by
the liver for disposal.

The liver synthesizes triglycerides and cholesterol, and packages them as

VLDLs before releasing them into the blood. When VLDLs (which consist
mainly of triglyceride) reach muscle and adipose blood vessels, their
triglycerides are hydrolyzed by LPL to fatty acids. The fatty acids that are
released are taken up by the surrounding muscle and adipose cells. During this
process, the VLDLs become progressively more dense and turn into LDLs.
While most of the resulting LDLs are taken up by the liver for disposal, some
circulate and distribute cholesterol to the rest of the body tissues.

HDLs, which are also secreted from the liver and intestine, have the task of
preventing lipid accumulation. They remove surplus cholesterol from tissues
and transfer it to LDLs that return it to the liver.

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Mobilization of triacylglycerol
The digestion of foodstuffs in and absorption
from the intestine characterizes the well-fed
state, which lasts for about four hours after a
meal. Free amino acids and glucose are
absorbed and transported to the liver. Excess
energy is converted to fat in the liver and
transported, along with dietary fat, to the
adipose tissues. The pancreas releases high
levels of insulin in response to these events.
Insulin signals the liver to convert glucose to
glycogen, amino acids to protein, and fat to
triglycerides. The adipose tissues synthesize
and deposit fats.
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As the individual enters the fasting state,
glycogen is broken down into glucose to supply
energy for the tissues. Simultaneously,
gluconeogenesis begins as amino acids,
lactate, and pyruvate from metabolism are
cycled into the formation of glucose. As fats
are broken down, the fatty acids supply energy
to the peripheral tissues, while the glycerol
from breaking down the triacylglycerols is
transported to the liver and converted to
glucose. Gluconeogenesis becomes more
important than glycogen breakdown after
about 16 hours of fasting.
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Ketone Bodies and Their Role in Metabolism

A special source of fuel and energy for certain tissues

 Some of the acetyl-CoA produced by fatty acid oxidation in

liver mitochondria is converted to acetone, acetoacetate
and b-hydroxybutyrate
 These are called "ketone bodies"
 Source of fuel for brain, heart and muscle
 Major energy source for brain during starvation
 In the event of low blood glucose, most other tissues have
additional energy sources besides ketone bodies (such as
fatty acids), but the brain does not
 Acetoacetate and β–hydroxybutyrate are transported from
the liver through the blood to target organs and tissues,
where they are converted to Acetyl-CoA.

Ketone Bodies and Diabetes

"Starvation of cells in the midst of plenty"

• Glucose is abundant in blood, but uptake by cells in muscle, liver, and

adipose cells is low
• Cells, metabolically starved, turn to gluconeogenesis and fat/protein
catabolism
• In type I diabetics, OAA is low, due to excess gluconeogenesis, so
Acetyl-CoA from fat/protein catabolism does not go to TCA, but rather
to ketone body production
• Acetone can be detected on breath of type I diabetics
• Diabetic Ketoacidosis results from a shortage of insulin; in response
the body switches to burning fatty acids and producing high level of
acidic ketone bodies.
• Vomiting, dehydration, deep gasping breathing, confusion and
occasionally coma are typical symptoms
• Untreated diabetic ketoacidosis can lead to death

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Words to know :
• Gluconeogenesis
• Fructose
– Polyol pathway
– Fructose intolerance (aldolase B deficiency)
• Galactose
– Lactose intolerance
– Galactosemia
• Glycogen
– Glycogen synthase
– Glucagon and Insulin
• Pentose phosphate pathway
– Glucose-6-P-Dehydrogenase Deficiency
• β-Oxydation
• Fatty acid synthesis

Next topics :

Amino acid metabolism

Nucleotide metabolism

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Reference

Moran, Horton, Scrimgeour, Perry, Rawn: Principles of Biochemistry,

Pearson New International Edition, 5th Edition, July 2013

End of Part II

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