Dr.

Joven Tanchuco, MD
2
OS 201
Correlative Human Cell Biology
Lec 8: Metabolic Regulation
– Proprionic acid (terminal 3-carbon
Outline molecule from beta oxidation of odd-
I. Regulation of Carbohydrate numbered fatty acids) may be converted
I. Regul Metabolism into dihydroxyacetone phosphate and
ation diphosphoglyceraldehyde, but there are
A. of Carbohydrates few odd-numbered fatty acids as sources
Carb 1. Characteristics • Amino acids are the major contributors in
ohydr glucose synthesis
ate  Available for the – May result in muscle wasting during
body in the form of starvation as muscles are mainly proteins
glucose
 Transported in the blood as glucose
 Stored as glycogen and triglycerides
 Only source of ATP when there is no oxygen
 Carbohydrates: up to 80-85% daily caloric
intake in less developed countries
 Lowest amount by weight (compared to CHON
and fats) in the body – about 2 kg in a 70 kg
person
2. Functions

a. Most important: energy production

 Polar  easily soluble in water, easily
transportable
 Easily diffused through cell membrane
(small molecule size overrides polarity,
presence of glucose transporters,
Figure 1. Overview of Carbohydrate Metabolism
concentration gradient after a meal)
 Abundance of CHO means most cells can
Notes:
use glucose as energy source in the post-
prandial stage (i.e. after a meal)
– Lactate formation has a limit because the
– Broken down in glycolysis
cell cannot tolerate high concentrations of
– Stored as glycogen or fatty acids
lactate as it may result in lactate acidosis;
b. Component in cell membranes as
lactate is converted back to glucose
glycoproteins
– Conversion of pyruvate to acetyl CoA is
 ex. ABO blood groups
one-way; acetyl CoA can’t be converted
c. Source of: oxaloacetic acid which replenishes
back to pyruvate, fat can’t be converted to
TCA cycle substrates, NADPH for fatty acid
carbohydrate
synthesis
Remember:
d. Provides precursor of glucoronic acid
e. Important component of nucleic acids (5
The liver is the only organ that will share its glycogen
carbon sugars, such as ribose, deoxyribose,
because it is the only organ with glucose-6-phosphotase.
are used for nucleic acid synthesis)
Glucose-6-phosphatase dephosphorylates glucose in the
f. Glucose in particular is the only major
liver when other tissues and organs are in need of it.
biomolecular energy source in the absence of
oxygen
 Prevents accumulation of NADH2 and • Glucose Homeostasis
FADH2 in their reduced forms • Many hormones in metabolic processes are
 2 ATPs produced in glycolysis may be life- controlled by glucose levels (ex. insulin, thyroid
saving for the cell (especially neurons hormone, epinephrine, glucagon)
which cannot regenrate) • Hypoglycemia is a more clinically dangerous
 Some cells can only use anaerobic situation because RBC, renal medulla cells,
glycolysis as its energy source (RBCs and nervous tissue rely almost exclusively on
which lack mitochondria) glucose for energy
B. Overview of Carbohydrate Metabolism • Insulin released when serum glucose levels go
1. Interconversion of Carbohydrates up (post-prandial state)
• Interconversion and intermittent feeding – Anabolic hormone thus stimulates
account for the fact that CHO are abundant in glucose entry and synthesis of glycogen,
the diet but only constitute a small amount in fatty acids, protein
cells (glucose  glycogen, fatty acids) – Inhibits lipolysis, beta oxidation,
gluconeogenesis
• Glucose must be maintained normally within a – Responds to serum glucose levels but
narrow concentration range thus other regulates also fatty acid and amino acid
biomolecules can also be converted to glucose metabolism
in between feedings (amino acids  glucose) – Works together with glucagons to
• Major glucose storage is in fatty acids regulate CHO metabolism
– Glycogen is more hydrated because it • As serum glucose and insulin levels drop:
remains polar thus its synthesis is limited – Glycogen breakdown and
by space requirements gluconeogenesis begin to maintain
– Fatty acids in the body are saturated and serum glucose levels
reduced and thus can be stored in a – Inhibition on lipolysis removed to allow
“dehydrated” form energy production from beta oxidation
• Fatty acid to glucose conversion is not possible

Anna, Everly, Edge, Julie Monday, Aug.24, 2009 Page 1 of 1

 Dr. Joven Tanchuco, MD
2
OS 201
Correlative Human Cell Biology
Lec 8: Metabolic Regulation
– Amino acid breakdown aids in Small intestine
gluconeogenesis and may act as
alternative energy source
C
2. Carbohydrate Absorption and
o
Transport  Need to transport against a gradient
t
• Carbohydrates in the diet are mostly from r
from lumen to epithelial cells. Kahit
plants and dairy products for lactose busog na ang cells mo with glucose,
a Na-
• Digestion starts in the mouth with salivary kailangan pa rin nya kuhanin ang
n depend
amylase though much of it occurs in the small glucose sa blood. 
s ent
intestine with pancreatic amylase and other p
dissacharides o
• Carbohydrates absorbed primarily as mono- r
Kidneys
and disaccharides t
• Absorption in the intestine takes place mainly  Glucose in the urine must be
via facilitated transport reabsorbed
– Co-transport with sodium Table 1. Different glucose transporters found inside the body
– Non-insulin dependent
– Further down the intestine absorption is • GLUT-4 stimulated by insulin  entry of
via active transport glucose into skeletal muscle and adipose is
• Entry into hepatic tissue (first organ insulin-dependent and occurs during high
encountered after entering portal circulation) is glucose levels
non-insulin dependent • Entry of glucose into pancreas, liver, and renal
TRANS- tubules is non-insulin dependent
LOCATION – Pancreas is responsible for secreting
PORTER
insulin
Ubiquitous, expressed to the largest
GLUT-1 – Renal tubule reabsorption of glucose
degree in brain and placenta must always be present especially in
β-cells in the pancreas, liver, kidney times of low serum glucose and insulin
– Liver is the first organ encountered and
cannot wait for an insulin signal
• Phosphorylation of glucose into glucose-6-
 High Km (15-20 mM)
phosphate inside the cell prevents its exit from
GLUT-2 the cell and decreases the amount of glucose
 Amount of glucose intake of these
organs are proportional to the amount within, allowing for more entry of glucose from
of glucose in the blood the blood
F – Glucose-6-phosphate may (1) enter
a pentose-phosphate pathway, (2) be
 Ensures that glucose is only taken
c used in glycogen synthesis, (3) undergo
up by the liver only when it is abundant
il glycolysis
GLUT-3 Ubiquitous
it C. Regulation of Glycolysis
a Adipose tissue, heart, skeletal muscle
1. Introduction
t GLYCOLYSIS
e
d • Major energy producing pathway in
 Only glucose transporter regulated
by INSULIN carbohydrate metabolism
D • Mainly a catabolic pathway but is also initial
if stage of glucose to fatty acid conversion
 Km is as high as the average
f
[serum glucose] which is 5mM • Major pathway for storage of energy
u
available in glucose
s
 When the [serum glucose] is high, • Glucose may also come from glycogen, not
i
GLUT-4 insulin causes translocation of these just the blood
o
transporters from an intracellular store • Only glycolytic reactions with the highest free
n
to the cell membrane energy change need to be regulated
because they are irreversible
 Fatty acids decrease the activity of REGULATION
GLUT-4 because it is the preferred
source of energy of cells 1. Regulation of glucose uptake through
glucose transporters (see table of
 In special situations: Anaerobic transporters above)
conditions, presence of respiratory  Glycolysis can be regulated through
poisons (e.g. Cyanide), uncoupling of the availability of the substrate
oxidative phosphorylation 2. Glycolysis can be regulated through the
GLUT-5 Small intestine control of specific reactions.
Rate-limiting Step: Characteristics

a. Small activity (small Vmax)

b. Small mass: action ratio
c. Products / reactants
d. Large negative free energy change.
(NOTE: this is the most important
criteria)

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 Dr. Joven Tanchuco, MD
2
OS 201
Correlative Human Cell Biology
Lec 8: Metabolic Regulation
There are 10 steps in the Glycolytic Pathway. Of these ten CoA for FA synthesis (both are
steps, three involve large amounts of negative free energy. stimulated by insulin)
These three steps are presented in the table below. – Liver thus undergoes little glycolysis as
it does not absorb much glucose,
Table 2. Three Reactions of Glycolysis with the Largest Negative sparing for other organs that need it
Free Energy • Catalyzed by hexokinase in non-hepatic tissue
Reacti – Low Km allows extraction of serum
Enzyme Description -∆G
on # glucose even in low concentrations
Phosphorylation -33.5 – Low Km also assures phosphorylation of
1 Hexokinase
glucose kj/mol glucose even if only small amounts are
Phosphofru- Phosphorylation of -22.2 taken into the cell
3
ctokinase fructose-6-phosphate kj/mol – At low insulin levels, glucose undergoes
Transfer of phosphate glycolysis; at high insulin levels, it is
Pyruvate -33.4
10 from phosphoenol- stored as glycogen and fatty acids
kinase kj/mol
pyruvate to ADP The signal for glycogenolysis in skeletal muscle is
These three reactions are catalyzed by enzymes whose epinephrine.
activities are irreversible. This is the primary reason why
they are potential sites of regulation in the glycolytic Always remember that the higher the Km value, the lower
pathway. the affinity of the enzyme for its substrate.

In general, insulin increases the transcription while

glucagon decreases the transcription of these three
enzymes. The effects of these hormones take place over a
period of hours to days and generally indicate whether a
person is well-fed or starving.

In general, if forward reaction is stimulated then backward

reaction is inhibited

2. Regulated Reactions
a. Glucose Glucose-6-phosphate

Hexokinase and Glucokinase

Figure 3. Km Value

b. Fructose-6-phosphate Fructose-1,6-
Table 3. Comparison of Glucokinase and Hexokinase phosphate
Glucokinase Hexokinase • Most critical for balance between
High Km (10 mM) Low Km (0.1 mM) gluconeogenesis and glycolysis
Inhibited by its product • Signal molecule for regulation is glucagons
Not inhibited by its product
(Glucose-6-phosphate) • Simultaneous stimulation of one direction
High Vmax Low Vmax and inhibition of the other  allows greater
Highly responsive in changes in flux and a more rapid response to
changes in glucose serum cell needs
concentration (more than – Greater substrates committed 
hexokinase) greater waste in overall effect 
Hepatic Tissues Extrahepatic Tissues greater response ability
Phosphorylation occurs Phosphorylation occurs – Ability to respond to changes more
only at high glucose levels even at low glucose levels important than baseline efficiency
Not regulated by insulin or
Insulin increases glucagon (the speed of its • Forward and backward reactions catalyzed
synthesis of this enzyme reaction makes it hard to by different enzymes: phosphofructokinase
regulate) (PFK1) for the forward and fructose-1,6-
Significance: Glucokinase in the liver has a higher Km biphosphatase (FBP) for the reverse
than hexokinase in extrahepatic tissues so that glucose Phosphofructokinase
will not be phosphorylated in the liver (it will not be trapped
in the liver). The high Vmax of glucokinase also allows the
liver to effectively store excess glucose circulating in the
blood. Phosphofructokinase catalyzes the rate-
limiting step in glycolysis and is the most
Elaboration: important control point.

• Catalyzed by glucokinase in the liver a. Allosterically inhibited by ATP

– Glucokinase has higher Km and is not
inhibited by glucose-6-phosphate thus
glucose is quickly metabolized in the
liver only when its levels are high
– High insulin levels mean that glucose is
stored as glycogen c/o glycogen
synthetase and fatty acids c/o malonyl
CoA synthetase which diverts acetyl
Anna, Everly, Edge, Julie Monday, Aug.24, 2009
meaning glycolysis is slowed when
cellular concentrations are high
b. Excess H+ inhibits glycolysis
because it favors the low-affinity state
of PFK  this helps minimize risk of
lactic acidosis
c. Citrate, an intermediate of the TCA
cycle, also inhibits PFK  signals
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Correlative Human Cell Biology
Lec 8: Metabolic Regulation
that biosynthetic precursors are
abundant
d. Allosterically activated by high levels
of AMP  AMP means that there is a
low source of ATP. When ATP has
already been used up, a high
concentration of ADP is generated.
From ADP, the cell can generate ATP
again through the reaction: ADP +
ADP  ATP + AMP, thus AMP is a
signal that ATP concentration is down
e. Allosterically activated by Fructose
2,6-bisposphate
f. Covalent regulation of PFK by
glucagon  high glucagon causes
phosphorylation of PFK2 which
catalyzes the forward reaction of
Fructose 6-Phosphate  Fructose
2,6-Phosphate (which we know can
activate PFK)
Remember: ATP, Citrate and H+ inhibits PFK. AMP
and Fructose 2,6-bisphosphate activates PFK.
• Bifunctional enzyme II has a kinase domain
(active when dephosphorylated) and a
phosphatase (active when
dephosphorylated)
– Phosphorylation regulated by blood
glucose level, mediated by
glucagon and insulin
– Fructose-6-phosphate can be
metabolized to fructose-2,6-
biphosphate c/o
phosphofructokinase 2 (PFK2)
– Fructose-2,6-biphosphatase (FBP2)
catalyzes conversion of fructose-
2,6-biphosphate to fructose-6-
phosphate
– Major regulator is glucagon
– Fructose-2,6-biphosphate
stimulates PFK1 (positive feedback)
and inhibits FBP (negative
feedback)
• Figure 4. Substrate Cycle as a Means of Control of
Glycolysis and Gluconeogenesis
Pyruvate Kinase
a. Activated by Fructose 1,6-bisphosphate
b. Allosterically inhibited by ATP and alanine
c. Covalently modified by glucagon (glucagons
causes phosphorylation which renders the
enzyme inactive)
Note:
Anna, Everly, Edge, Julie Monday, Aug.24, 2009
Dr. Joven Tanchuco, MD
2
1. Kinases and phosphatases are different domains
of one bifunctional polypeptide enzyme.
2. The substrate cycle involving Fructose 2,6-
bisphosphate is a potent regulator of both
glycolysis and gluconeogenesis
3. This ensures that gluconeogenesis and glycolysis
are not occurring simultaneously in order to avoid
a futile cycle
4. Fructose 2,6-bisphosphate is the substrate that
has the regulatory effect
a. Most potent activator of PFK I
b. Inhibitor of Fructose 1,6-phosphatase
Phosphofructokinase II (PFK II)
• In the unphosphoryated state, has kinase
function
• In the phosphorylated state, has phosphatase
function
• Part of covalent modification  inhibits fructose
biphosphate and stimulates fructokinase
Glucagon
• Hormone which is produced when blood glucose
is low
• Stimulates gluconeogenesis
• binds to plasma membrane receptors on liver
cells, activating membrane-localized adenylate
cyclase, leading to an increase in the conversion
of ATP to cAMP
Recall: cAMP  protein kinase b  protein kinase a
• Protein kinase phosphorylates F 2,6-phosphatase
(FBP II) and inhibits PFK II, leading to a decrease
in F 2,6-biphosphate
• No more inhibition of F 1,6-bis pase (FBP), which
dephosphorylates F 1,6 biphosphate to F 6
phosphate
• Fructose 6 phosphate is converted to glucose
(a.k.a. GLUCONEOGENESIS)
Insulin
• opposes all effects of glucagons
• leads to the dephosphorylation of PFK II (active)
which will eventually stimulate FPK through F
2,6-biphosphate
• also leads to a decrease of cAMP
Allosteric modification of FBP
• high amounts of AMP: stimulates PFK
– inhibits the production of glucose
through gluconeogenesis and favors the
production of ATP (energy) through
glycolysis
• high amounts of ATP, citrate and H+: inhibits PFK
– breakdown of glucose through glycolysis
is no longer needed
Feedforward Stimulation
• low fructose-2,6-biphosphate, high fructose-
6-phosphate  gluconeogenesis
Glucagon, acting on adenyl cyclase receptor complex
and through a protein kinase, can inhibit PFK2 and
stimulate FBP2
– Low glucose high glucagonlow
fructose-2,6-biphosphatehigh
fructose-6-phosphate
– Flux is towards gluconeogenensis
because of lack of inhibition of FBP by
fructose-2,6-biphosphate
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Lec 8: Metabolic Regulation
• high fructose-2,6-biphosphate, high fructose
-1,6-biphosphate  glycolysis
Low glucagon (high blood sugar) 
dephosphorylation of the enzyme II conversion to
fructose-2,6-biphosphate from fructose-6-phosphate
 stimulatory effect on PFK1 glycolysis
Phosphoenol pyruvate pyruvate
• Amino acids as the source of pyruvate 
prevents futility of the process
• Includes the Malate shuttle (malate exits
mitochondrion)
• Will happen simultaneous with the breakdown of
carbohydrates
• Strictly irreversible
• Last reaction in glycolysis which is regulated.
• Catalyzed by pyruvate kinase and produces 1
ATP
Figure 5. The Pyruvate-Phosphoenol Pseudocycle
a. Phosphoenol pyruvate converted to pyruvate via
pyruvate kinase
○ This enzyme is stimulated by fructose-
1,6-biphosphate and inhibited by ATP,
glucagon, alanine, and phenylalanine
(these 2 amino acids are substrates for
gluconeogenesis)
○ Inhibition achieved through covalent
modification with the phosphorylated
pyruvate kinase being the inactive form
○ Alanine makes pyruvate kinase more
susceptible to covalent modification
○ PEPpyruvate cycle produces 1 ATP
b. Pyruvate formed enters mitochondrion to be
converted to acetyl CoA c/o pyruvate
dehydrogenase (PDH), another strictly
irreversible reaction
○ Reason why fatty acids cannot be
converted to glucose
○ Acetyl CoA from fatty acid breakdown
cannot regenerate pyruvate and instead
Anna, Everly, Edge, Julie Monday, Aug.24, 2009
Dr. Joven Tanchuco, MD
2
enter the TCA cycle where its 2 carbons
are released as CO2
○ Enzyme also regulated by covalent
modification
 Also active in the
dephosphorylated form c/o
PDH phosphatase which is
stimulated by insulin
 Inactivation via PDH kinase
which is stimulated by acetyl
CoA, NADH, ATP
 Pyruvate inhibits PDH kinase
and activates PDH
 Dephosphorylation of PDH is
done by specific PDH
phosphatase
 PDH phosphatase is stimulated
by insulin
 Insulin promotes conversion of
pyruvate to acetyl CoA
c. Pyruvate can be converted to oxaloacetate via
pyruvate carboxylase
○ Stimulated by low glucose levels  low
insulin  PDH becomes inactive
○ This allows conversion of pyruvate to
oxaloacetate by pyruvate carboxylase
○ Pyruvate carboxylase is stimulated by
acetyl CoA from beta oxidation of fatty
acids when serum glucose and insulin
levels are low
○ Pyruvate carboxylase is however
inhibited by ADP
○ The reaction initiates regeneration of
phosphoenol pyruvate from pyruvate
d. Oxaloacetate is converted to malate via malate
dehydrogenase
○ Oxaloacetate is converted to malate
inside the mitochondrion by malate
dehydrogenase
○ Malate is transported out of the
mitochondrion into the cytosol (malate
shuttle)
○ Oxaloacetate regenerated from malate
via same enzyme (malate
dehydrogenase)
○ Oxaloacetate converted to phosphoenol
pyruvate via PEP carboxykinase
○ Synthesis of PEP carboxykinase is
stimulated by glucagon in the presence
of glucocorticoids
 Inhibited by ADP, insulin and
high level of glucose
e. Pyruvate can be converted to acetyl coA by
pyruvate dehydrogenase.
○ Pyruvate is transferred to mitochondrion
○ Converted to acetyl CoA by pyruvate
dehydrogenase (irreversible reaction)
○ Acetyl CoA enters TCA
○ Carbons are released as CO2
• The whole pyruvatePEP cycle consumes 2
ATPs equivalents
Page 5 of 5
 Dr. Joven Tanchuco, MD
2
OS 201
Correlative Human Cell Biology
Lec 8: Metabolic Regulation
• The reverse process of the PEP pyruvate ○ Blood-borne glucose is converted by
produces only 1 ATP (there is a net requirement exercising muscle to lactate which
of another unit of ATP) diffuses into the blood. This lactate is
taken up by the liver and converted to
• Insulin and glucagon also regulate protein glucose which is released back to the
circulation or returns to the muscle to
synthesis and amino acid breakdown,
replenish their glycogen.
respectively
a. Promote gluconeogenesis and amino ○ 2 lactate  2 pyruvate  glucose
acid breakdown (Liver)  glucose  2 pyruvate  2
lactate (muscle)
b. High glucagon/insulin ratio  promotes ○ Significance: Cramps is a signal for the
synthesis of PEP carboxykinase,
glucose-6-phosphatase, other body to stop using energy. Too much
aminotransferases lactate is being produced.
c. inhibits glucokinase synthesis and b. Glucose alanine cycle
pyruvate kinase
○ Amino acids in the muscle are converted
to alanine (small molecules) which are
3. Gluconeogenesis easily passed on to the liver to create
new glucose
 Formation of glucose from nonhexose
precursor
○ Nitrogen (carried by alanine) is
 Gluconeogenesis is NOT the reverse eliminated through the urea cycle which
of glycolysis and vice versa. occurs only in the liver
○ Alpha keto acid is used for energy
○ Glucose can be exported from the liver
to the other cells.
Glycolysis Gluconeogenesis ○ Gluconeogenesis in the liver allows AA
Glucose to Pyruvate to glucose BUT to be broken down in the muscle.
pyruvate with detour
Glucose = Glucose = product
substrate c. Entry of glycerol derived from triglycerides into
Product: 2 Substrate: amino acids gluconeogenesis
molecules of (ketoacids)
pyruvate, ATP and ○ Glycerol is converted to dihydroxyacetone
NADH phosphate (DHAP), which could then be
converted to glucose
Makes 2 moles of Consumes 6 ATP
ATP  Regulation of glycogen metabolism

○ Controlled by the regulation of key enzymes

 Glucagon – main controlling hormone
 Fructose-6-phosphate:fructose-1,6-biphosphate
substrate cycle = determines flux between
glycolysis and gluconeogenesis
 Release of glucose from inside the cell would be
made possible with the removal of phosphate
done by glucose -6-phosphate (found only in liver
and renal cells)
 Reactions converting amino acids into glucose
depends on type of amino acid and their
corresponding ketoacids
 About 150 grams of glucose per day can be
made in gluconeogenesis.
 Only liver and kidney are capable of
gluconeogenesis.
 Glucagon - major hormone controlling
gluconeogenesis
 New glucose come from:
○ Breakdown of AA except leucine and
lysine (pyruvate)
○ Lactate (Cori cycle)
○ Glycerol backbone of TAG
○ Propionyl CoA (succinyl CoA)
 Regeneration of glucose
○ Glucose can be regenerated in the liver from
lactate and alanine in the muscle.
a. Cori Cycle (lactate)
○ Glucose is regenerated in the liver from
lactate
glycogen phosphorylase (involved in
glycogenolysis) and glycogen synthetase
(involved in glycogenesis)
○ Glycogen synthesis and breakdown occurs in
cytosol.
○ Glycogenolysis – stimulated by high levels of
AMP and inorganic phosphate (Pi) and
inhibited by ATP and glucose-6-phosphate
○ To prevent futile cycles, simultaneous
regulation of both pathways has to be present:
○ Glycogen phosphorylase
• facilitates conversion of glycogen to
glucose
• stimulated by high levels of AMP and
Pi;
• inhibited by ATP and glucose-6-
phosphate
• Active enzyme is independent of these
but requires Ca2+
• Epinephrine in muscle and glucagon in
liver activates the enzyme
• Less sensitive to allosteric modifier
○ Glycogen synthase
• Involve in glycogen synthesis
• Facilitates conversion of glucose to
glycogen
• Stimulated by insulin
• allosterically actived by glucose-6-
phosphate, a precursor of glucose-1-

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Correlative Human Cell Biology
Lec 8: Metabolic Regulation
phosphate, the building block of
glycogen.
• Has seven sites of phosphorylation
catalyzed by seven protein kinase
• Phosphorylation of these sites lead to
progressive inactivated of the enzyme
• Encourages phosphorylation
• is a covalent modification of glycogen
phosphorylase
Figure 6. Covalent Modification of the glycogen
phosphorylase enzyme. Glycogen Phosphorylase is activated
by the linkage of a phosphate prosthetic group. This is
catalyzed by phosphorylase kinase which is also activated by
phosphorylation of catalyzed by another protein kinase.
Activation of protein kinase b is done by presence of high
levels of cAMP, formed from ATP by adenyl cyclase located in
the cell membrane and associated by cell membrane
receptors. In glycogen phosphorylase enzyme system, cell
receptor is specific for glucagon (glycogenolysis in liver) or
epinephrine (glycogenolysis in skeletal muscle). The round-
about way of activating glycogen phosphorylase with a signal
from glucagon or epinephrine is called amplification.
✔ The difference in signal molecule illustrates basic
difference in the functions of major tissue involved
in glycogenolysis.
✔ Skeletal muscles involved primarily in movement
would need more energy.
✔ Epinephrine would allow glycogenolysis to take
place to provide lot of glucose needed for rapid
production of energy.
 Regulation of Glycogen Synthesis and
Degradation
○ Regulation of both glycogen synthesis and
degradation are important in maintaining blood
glucose levels
○ Liver
• Glycogen synthesis = occurs during
well-fed state
• Glycogen degradation = occrs during
periods of fasting
○ Muscle
• Glycogen synthesis = occurs during
active exercise
• Glycogen degradation = occurs as soon
as muscle is again at rest\
○ Regulation occurs and is accomplished via
glycogen synthase and glycogen phosphorylase
on 2 levels:
• Allosteric- allosteric enzymes having
different conformation induced by the
binding of modulators
Anna, Everly, Edge, Julie Monday, Aug.24, 2009
Dr. Joven Tanchuco, MD
2
• Hormonal - presence of insulin,
glucagon, fight/flight hormones)
[the following are from 2013 trans ]
 Allosteric Regulation of glycogen synthesis and
degradation
○ Glycogen synthase and glycogen phosphorylase
respond to levels of:
• metabolites / substrates
• energy needs
○ Glycogen synthesis = stimulated when substrate
availability and energy levels are high
○ Glycogen degradation = stimulated when energy
levels and available glucose are low
a. Regulation in the well-fed state
• Glycogen synthase: allosterically
activated by glucose 6-phosphate (G6P)
• Glycogen phosphatase: allosterically
inhibited by G6P, ATP, and in the liver,
glucose
b. Regulation in muscle by calcium
b.1. In muscle contraction: rapid and urgent need
for ATP
– Nerve impulses cause membrane
depolarization  Ca2+ release from
sarcoplasmic reticulum into sarcoplasm
of muscle cells
– Ca2+ binds to calmodulin
– Binding of 4 molecules of Ca2+ triggers a
conformational change = Ca2+ -
calmodulin complex
– Ca2+ - calmodulin complex binds to and
activates phosphorylase kinase (without
phosphorylation)
b.2. in muscle relaxation
– Ca2+ returns to the sarcomplamsic
reticulum  phosphokinase becomes
inactive
c. Regulation in muscle by AMP
• Glycogen phosphatase is active in the
presence of high AMP concentrations (e.g.
during anoxia and ATP depletion)
• AMP binds to the inactive form of glycogen
phosphatase  activated (without
phosphorylation)
 Activation of glycogen degradation by cAMP-
directed pathway
○ Binding of hormones (e.g. glucagon or
epinephrine) to membrane receptors signals
the need for glycogen breakdown to:
• elevate blood glucose levels
• provide energy for exercising muscle
a. activation of protein kinase
• binding of hormones to receptors
cAMP-mediated activation of cAMP-
dependent protein kinase
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Lec 8: Metabolic Regulation
• protein kinase: tetramer
– 2 regulatory subunits
– 2 catalytic subunits
• cAMP binds to the regulatory
subunit dimer  release of
individual catalytic subunits (active)
b. Activation of phosporylase kinase
• Active cAMP-dependent protein
kinase phosphorylates inactive
phosphorylase kinase  activation
C. Activation of glycogen phosphatase
• Glycogen phosphatase exists in 2
forms:
– Inactive “b” form (GP B)
– Active “a” form (GP A)
• Active phosphokinase phosphorylates
GP B to active GP A
• GP A begins glycogen breakdown
• Reconversion of GP A to GP B:
hydrolysis of its phosphate by protein
phosphatase 1
• AMPLIFICATION of hormonal signal
 Inhibition of glycogen synthesis by cAMP-directed
pathway
• Glycogen synthase: regulated enzyme in
glycogen synthesis
• 2 forms:
a) a form: not phosphorylated, most active
b) b form: phosphorylated, inactive
• glycogen synthase is converted to b form by
phosphorylations at a # of sites  catalyzed
by a cAMP-dependent protein kinase
• protein kinase phosphorylates and thereby
inactivates glycogen synthase
4. Pentose-Phosphate Pathway
*He skipped this, but it was part of his PowerPoint
presentation and was also in the 2013 trans so...
 Also called the hexose-monophosphate shunt or
6-phosphogluconate pathway
 Occurs in the cytosol
 Has two irreversible oxidative reactions:
A. Dehydrogenation of glucose 6-phosphate
○ Glucose 6-phosphate dehydrogenase
(G6PD) is strongly inhibited by NADPH
○ Insulin enhances G6PD gene expression
○ Flux through the pathway increases in the well-
fed state
○ Produces first NADPH
B. Formation of ribulose 5-phosphate
○ Produces second NADPH and 1 CO2
 Reversible nonoxidative reactions (sugar–
phosphate interconversions)
○ Permit ribulose 5-phosphate to be converted to
either:
○ Ribose 5-phosphate
– When demand for ribose for incorporation
into nucleotides and nucleic acids is greater
than for NADPH
Anna, Everly, Edge, Julie Monday, Aug.24, 2009
Dr. Joven Tanchuco, MD
2
– When G6PD is inhibited, ribose can be
synthesized glyceraldehyde 3-phosphate
and fructose 6-phosphate
– When more ribose is needed, ribose may be
regenerated anew from glucose 6-phosphate
from exogenous glucose
 Intermediates of glycolysis (fructose 6-
phosphate and glyceraldehyde 3-
phosphate)
– When demand for NADPH is greater than for
ribose 5-phosphate (for reductive
biosynthetic reactions)
• No ATP is directly consumed or produced
• Products
○ CO2 from carbon 1 of glucose 6-phosphate
○ 2 NADPH per glucose 6-phosphate entering
the oxidative part of the pathway
– Provides a major portion of the
body’s NADPH
– NADPH: biochemical reductant;
needed in fatty acid synthesis
○ Ribose 5-phosphate
– For biosynthesis of nucleotides
• Found in tissues active in reductive biosynthesis:
Liver, adipose, adrenal cortex, gonads,
erythrocytes
IV. Regulation of Fatty Acid and Cholesterol
Metabolism
A.Introduction
 Fatty acids and cholesterol belong to the lipid group
of biomolecules. They are the best way for storing
energy in the cell.
 Most of the lipids found in humans are in the form of
triglycerides.
a. One of the most efficient forms of storing energy
b. Contain six times the amount of calories on a
weight basis as compared to carbohydrates
c. Less oxidized (contains less oxygen per carbon
atom)
d.Less hydrolysed because they are non-polar 
allow more oxidation to take place, stored in
compact form without much water.
e. Body stores 80% of the energy available to it as
fats.
B.Overview of Lipid Metabolism
Figure 7. Overview of Fatty Acid Metabolism
 The hydrophobic property of fatty acids allow them
to pass easily across the cell and organell
membranes.
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 Dr. Joven Tanchuco, MD
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OS 201
Correlative Human Cell Biology
Lec 8: Metabolic Regulation
 To make the fatty acids impermeable, that is to trap
them and make them commit to a metabolic
pathway, they are combined with a large molecule,
the Coenzyme-A (CoA) by the action of the enzyme
Acyl CoA synthetase.
 However, beta-oxidation occurs in the inner
mitochondrial matrix. The enzyme Carnitine
Palmitoyl Transferase (CPT-1) transports the fatty
Acyl CoA from the cytosol into the inner
mitochondrial matrix.
 Once inside the mitochondrial matrix, the FA is
already committed to take the beta-oxidation
pathway.
 Fatty acid from the diet is converted to its storage
form, the triglycerides through esterification.
 Ketogenesis in the liver occurs when TCA
intermediates are inadequate (e.g. low [glucose]) to
allow complete oxidation of FAs. This allows the
energy of the FAs to be extracted and redistributed Figure 8. Lipolysis and Esterification. Encircled portion indicates
in the form of more soluble ketone bodies. pathway that proceeds when there is low blood glucose. Solid
arrows-activation; broken arrows-inhibition.
 Fatty acids are also derived from glucose and
amino acids  even if fat intake is reduced, excess c. High [glucose]  high [insulin]  TAGs from the
calories from CHOs are stored as fats. intestines must gain entry into adipose cells for
C.Lipolysis and Esterification storage  lipoprotein lipase is active.
 Esterification – FA molecules are added to the d.Low [glucose]  low [insulin]  there is no
glycerol backbone; esterase enzyme inhibition of the hormone-sensitive lipase (it is
 Lipolysis – hydrolytic removal of the FA moiety from active)  breakdown of TAGs in the adipose cell
the glycerol backbone of triglycerides; lipase  FFA made available to other cells as a source
enzyme. of energy.
a. Lipoprotein lipase – located on the endothelial
surface of blood vessels; breaks up TAGs coming Table 1. Hormonal regulation of the Hormone-sensitive and
from the intestines; allow entry of FAs into the Lipoprotein lipase enzymes.
adipose cells. Enzyme Regulatory Agent Effect
b. Hormone sensitive lipase – located in the cytosol;
breaks up TAGs stored inside the adipose cells. Hormone Lipolytic hormones Stimulation by cAMP-
insulin
Sensitive (epinephrine, mediated
Lipase glucagons, ACTH) phosphorylation of
relatively inactive
Insulin enzyme

Prostaglandin Inhibition

Inhibition
Lipoprotein Lipoprotein Activation
Lipase
Apoprotein C-II

Insulin Activation

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 Dr. Joven Tanchuco, MD
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Correlative Human Cell Biology
Lec 8: Metabolic Regulation
the needed TCA intermediates to complete the
oxidation of FA.
 There is now a difficulty occurring in the TCA cycle
because there is no enough intermediates to
process the acetyl-CoA coming in.
 A pseudo high-energy state is created because of
the accumulation of acetyl CoA (as what happens
during starvation)
 Ketogenesis relieves the TCA cycle of the stress as
2 molecules of Acetyl-CoA form acetoacetate 
inhibited accumulation.
 Ketone bodies are also more soluble and easily
transported.
 Occurs only in the liver because of the high levels
of activity of the enzymes HMG-CoA synthase and
HMG-CoA lyase.
 Factors affecting Ketogenesis:
a.High lipolysis  serum [FFA]  liver is more
saturated  high ketogenesis
b.High insulin/glucagons ratio  (+) acetyl CoA
Figure 9. Covalent modification of hormone-sensitive lipase. carboxylase  high [malonyl CoA]  (-) CPT-1
 decreased low ketogenesis. (Activity of CPT-1
D.Beta-oxidation determines whether the initial steps of b-oxidn
and ketogenesis takes place or not)
c. Gluconeogenesis in the liver also utilizes
oxaloacetate  lesser ability of the TCA cycle 
increased ketogenesis
 Ketoacidosis – results when there is excessive
ketogenesis; seen in people with diabetes mellitus
or under starvation

Figure 10. Interrelationships between glycolysis and fatty acid

metabolism. Solid circle-activation; hollow circle-inhibition.

 CPT-1 is inhibited by malonyl coA, an intermediate

of FA sythesis.
 Insulin indirectly inhibits CPT-1 by activating acetyl Figure 11. Ketogenesis
CoA carboxylase which in turn converts more acetyl F. Lipogenesis
CoA to malonyl coA.  High in people with high carbohydrate intake
 Acetyl CoA carboxylase is the committed enzyme in  Low in people with restricted caloric intake, high fat
FA synthesis. diet and low insulin levels
 Acetyl CoA, which needs to be recycled, is
transported out of the mitochondrion through the
citrate shuttle.
 FFA (palmitic acid in the diagram) serves as a
signal that there are adequate amounts of FA thus
inhibiting FA synthesis through acetyl CoA
carboxylase.
 During fasting, glucose is converted to energy and
not to FA.
 Too much FFA:
a. Tissue would prefer FFA, especially skeletal
muscle.
b. Glucose will be spared.
c. Cells will not need glucose
d. Glucose uptake will not be significant.
e. Insulin levels could remain high
(Hyperinsulinemia).
E.Ketogenesis
 FA break down occurs in conditions of relatively low
glucose. However, glucose is needed to generate

Anna, Everly, Edge, Julie Monday, Aug.24, 2009 Page 10 of 10

 Dr. Joven Tanchuco, MD
2
OS 201
Correlative Human Cell Biology
Lec 8: Metabolic Regulation
 Hyperinsulinemia  Hypercholesterolemia

Julie: Hi Dana and Laureen! Hi Psych my loves! Hi Bryan,

Ricky and Joreb! Thanks for keeping me company; no
thanks for making me watch that stupid slasher movie. Hi
Dior, Chax, Nino, JB Boy, Jana, Ruby, Shasha, kung
sinuman sa inyo ang maging buddy ko!

Anna: Kamusta naman? Siguro ay nakangiti ka habang

binabasa mo ito, nagbabakasakali na ikaw ay mababati..
hehe. Kaya para naman ikaw ay masiyahan, kahit sino ka
man, buong galak kitang binabati sa trans na ito. AJA!
(>’-’)> sa mga behsci pips -robertojegargracestephrenz
-tandaan nyo ang theme song natin.. ang walang
kamatayan (If we hold on together!) ahaha.. pipahatid ko
din ang pagbati sa masiyahing tao sa kabilang pampang –
jeshjaybeadavidalphiusms.drogerlance – kaway naman
jan! sa mga masisipag na rso, carry on! at sa aking
magigiting na seatmates, tulugan na! 

Figure 12. Lipogenesis and Glycolysis. Everly: Sorry sa mahabang trans. Kasalanan ni ed! Hello
 Main regulatory points: to the IB people especially to Bea, Marvin and Al.
a. Acetyl CoA carboxylase Godbless sa metab exam natin.
○ Stimulation by citrate indicates wide availability
of glucose and raw materials for FA synthesis
○ Inhibited by acyl CoA means that FAs are
coming in from the diet
b. Pyruvate dehydrogenase

○ Inhibited by high [acetyl-CoA], [NADH] and

[ATP] through covalent modification; indicates
a high-energy state.
G.Cholesterol Metabolism

Figure 13. Regulation of Cholesterol Metabolism.

 Negative feedback mechanism: cholesterol inhibits
HMG CoA reductase by inhibition of the enzyme
synthesis or activation of the enzymes that degrade
HMG CoA reductase.
 Activation by insulin is through covalent
modification.
 HMG CoA reductase is active at the
dephosphorylated state.
 Level of cholesterol in the cell:
a.Increased uptake by the number of LDL
receptors  inhibition of cholesterol synthesis
b. Cholesterol inside the cell downregulates number
of LDL receptors.
c. Decreased by loss of cholesterol from cell
membranes as HDL, esterification and synthesis
into cholesterol derivatives.
Anna, Everly, Edge, Julie Monday, Aug.24, 2009 Page 11 of 11