Metabolic Integration in the

Fed and Starve States

Endocrine Module – Med 7524

(Biochemistry)
 Learning competencies

1.Discuss the mechanisms that control the flow of

metabolites through metabolic pathways
2.Explain the metabolic role of the liver, adipose
tissue, skeletal muscles and brain in the well-
fed and fasting state
3.Discuss the intertissue relationships in the
starve-feed cycle
4.Compare the metabolic effects of insulin and
glucagon
 Four mechanisms that control the flow of
metabolites through metabolic pathways

◼ Availability of substrates

◼ Allosteric activation/inhibition of enzymes

◼ Covalent modification of enzymes

◼ Induction-repression of enzyme synthesis

 Time scale in which each mechanism
takes effect

◼ Substrate availability – within minutes

◼ Allosteric effect on enzymes – within minutes

◼ Covalent modification of enzymes – minutes

to hours

◼ Enzyme synthesis – hours to days

 Allosteric Effects

◼ Involve rate-limiting enzymes

◼ They give a sigmoid curve when concentration
of the enzyme is plotted vs reaction velocity
◼ Ex., Effects of fructose 2,6-bisphosphate

1) Activates phosphofructokinase →stimulation

of glycolysis

2) Inhibits fructose 1,6-bisphosphatase →

inhibition of gluconeogenesis
 Covalent Modification of Enzymes

◼ Involves addition or removal of phosphate

from ser, thr and tyr residues

◼ Ex., Effects of phosphorylation by kinase

◼ 1) Activation of glycogen phosphorylase

◼ 2) Inactivation of glycogen synthase

Induction/Repression of Enzyme Synthesis

◼ Enzymes involved are those that are needed

under selected physiologic conditions

◼ Example: in the fed state, insulin promote

synthesis of key enzymes involved in:

1. lipogenesis and triglyceride synthesis

2. DNA, RNA and protein synthesis
METABOLISM IN THE WELL-FED
(ABSORPTIVE) STATE
METABOLISM IN THE WELL-FED (ABSORPTIVE) STATE

LIVER: THE MAJOR PROCESSING

ORGAN OF THE BODY
 Role of the liver in carbohydrate
metabolism
◼ More of a glucose-producer during the day
except after meals
◼ Retains 60% of glucose from the diet

◼ Increased glucose metabolism is due to:

1. Increased formation of glucose 6-phosphate

by glucokinase (high Km for glucose)

2. Increased glycogenesis because of activation

of glycogen synthase
 Liver in CHO Metabolism
◼ 3. Increased HMP shunt due to:
- availability of glucose 6-PO4
- increased activity of glucose 6-PO4 DH
- increased utilization of NADPH for
hepatic fatty acid synthesis

◼ 4. Increased glycolysis due to

- activation of phosphofructokinase and
pyruvate kinase
- glucose is converted to acetyl CoA
- acetyl CoA → Krebs or lipogenesis
 Liver in CHO Metabolism

◼ 5. Decreased gluconeogenesis due to

- inactive pyruvate carboxylase 2o to
low levels of acetyl CoA

- high insulin:glucagon ratio leads to

inactivation or repression of:
a) phosphoenolpyruvate carboxykinase
(PEPCK)
b) fructose 1,6-bisphosphatase
 Role of the Liver in Fat Metabolism
◼ 1. Increased lipogenesis due to availability
of acetyl CoA and NADPH
- activation of acetyl CoA carboxylase

◼ 2. Increased TAG synthesis due to increased

availability of fatty acids
* Fatty acids are derived from:
a) lipogenesis
b) hydrolysis of TAG in chylomicron remnant

* Availability of glycerol 3-PO4 from glycolysis

 Role of the Liver in Protein Metabolism
◼ 1. Increased amino acid degradation due to
excess amino acids from the diet.
* AA are deaminated forming their carbon
skeletons (α-keto acids)
- exempted are the branched-chain AA

◼ 2. Release of surplus amino acids into the

blood

◼ 3. Increased protein synthesis

METABOLISM IN THE WELL-FED (ABSORPTIVE) STATE

ADIPOSE TISSUE: ENERGY STORAGE

SITE OF TRIACYLGLYCEROL
 Role of the Adipose Tissue in CHO
Metabolism

◼ 1. Increased glucose transport into fat cells

- due to sensitivity to insulin

◼ 2. Increased glycolysis due to increased

availability of glucose from the diet
- Glycolysis supply DHAP which is converted
to glycerol 3-PO4 for TAG synthesis

◼ 3. Increased activity of HMP shunt → NADPH →

inc. lipogenesis → inc. TAG synthesis
 Role of the Adipose Tissue in Fat
Metabolism

◼ 1. Increased lipogenesis
- de novo synthesis of fatty acid is not a
major pathway

◼ 2. Increased TAG synthesis due to:

a) release of fatty acid from hydrolysis of
TAGs in chylomicron and VLDL
catalyzed by lipoprotein lipase
b) availability of glycerol 3-PO4 coming
from DHAP (glycolysis)
 Adipose tissue in Fat Metabolism

◼ 3. Decreased lipolysis due to inactivation of

hormone-sensitive lipase

- insulin favors the dephosphorylated,

inactive state of lipase
METABOLISM IN THE WELL-FED (ABSORPTIVE) STATE

SKELETAL MUSCLE: AN
OXIDATIVE TISSUE

Oxygen consumption:
- 30% at rest
- 90% during strenuous exercise
 Role of the Skeletal Muscle in CHO
Metabolism

◼ 1. Increased glucose transport into muscle

- major source of fuel of resting muscle
◼ 2. Increased glycolysis

◼ 3. Increased glycogenesis due to increased

availability of glucose 6-PO4

- results from increased insulin:glucagon

ratio
 Role of the Skeletal Muscle in Fat and
Protein Metabolism

◼ Lipoprotein lipase releases fatty acids from

* chylomicrons and VLDL
◼ Fatty acids are only of secondary importance
as a fuel for muscle in the absorptive
state

- Increased uptake of amino acids and protein

synthesis
- Increased storage of ATP
- Increased storage of creatine phosphate
METABOLISM IN THE WELL-FED (ABSORPTIVE) STATE

BRAIN: ENERGY CONSUMER

RATHER THAN ENERGY GIVER

Accounts for 20% of oxygen

consumption of the body at rest
 Role of the Brain in CHO Metabolism

◼ Uses glucose exclusively as a fuel in the

absorptive state

◼ Dependent on the supply of blood glucose

◼ Oxidizes 140 g/day of glucose

◼ Does not store glycogen

 Role of the Brain in Fat Metabolism

◼ Does not store TAG

◼ Fatty acids do not sufficiently cross the

blood-brain barrier

◼ Does not use fatty acids for its energy needs

 METABOLISM IN THE
STARVE (FASTING) STATE
 Starvation
◼ Causes:
1. inability to obtain food
2. inability to eat because of disease
3. desire to lose weight rapidly

◼ During this catabolic periods there are

exchanges of substrate among organs

◼ There are two priorities of metabolism:

1. maintain adequate plasma levels of
glucose for use by the brain
2. mobilize fatty acids and ketone bodies
to supply energy to other tissues
The Role of the Liver in CHO Metabolism

◼ 1. Increased glycogenolysis
- due to phosphorylation and activation
of glycogen phosphorylase
- glycogen store is nearly depleted after
10 to 18 hours of fasting

◼ 2. Increased gluconeogenesis
- substrates used are amino acids, glycerol
and lactate
- starts 4 to 6 hours after the last meal
- maintains blood glucose during overnight
and prolonged fasting
The Role of the Liver in Fat Metabolism

◼ 1. Increased beta oxidation

- fatty acids obtained from lipolysis in
adipose tissue

◼ 2. Increased ketogenesis due to excess

acetyl CoA obtained from beta oxidation

- starts during the first days of starvation

- use for energy by the brain, heart, etc.
- slows down the loss of tissue protein
- reduces the need for gluconeogenesis
 The Role of Adipose Tissue in
Starvation
◼ 1. Carbohydrate metabolism
- decreased transport of glucose into fat cells
- decreased lipogenesis and TAG synthesis

◼ 2. Fat metabolism
a) Increased lipolysis due to phosphorylation
and activation of hormone-sensitive lipase
by catecholamines
- increased release of fatty acids

b) Decreased uptake of fatty acids due to

low activity of lipoprotein lipase
 Skeletal Muscle in Starvation

◼ 1. Carbohydrate metabolism
- decreased glucose transport into muscles
- due to decreased insulin:glucagon ratio

◼ 2. Lipid metabolism
- first 2 weeks, muscle uses fatty acids and
ketone bodies
- in 3 weeks, oxidizes fatty acids almost
exclusively
 Skeletal Muscle in Starvation

◼ 3. Protein metabolism
- first few days, there is rapid proteolysis
- amino acids are used for gluconeogenesis

- alanine and glutamine are the most

important gluconeogenic amino acids

- after several weeks, there is decreased

muscle proteolysis
- due to decline in the need for glucose
as source of energy for the brain
 The Brain during Starvation

◼ First days of starvation (early fasting), it

uses glucose exclusively

◼ In prolonged starvation (> 2 to 3 weeks of

fasting), it uses ketone bodies as fuel

*Utilization of ketone bodies reduce need for

protein catabolism for gluconeogenesis
 Metabolic interrelationships during the
early fasting state

◼ Hepatic glycogenolysis is the most important

source of blood glucose

◼ Cori cycle become active

- conversion of glucose to lactate (glycolysis) in
the peripheral tissues followed by conversion of
lactate back to glucose (gluconeogenesis) in the
liver

◼ Glucose-alanine cycle become active

 Metabolic interrelationships during the
fasting state
◼ Increased hepatic gluconeogenesis
◼ Substrates: lactate, glycerol and alanine

◼ Fatty acids cannot be converted to glucose

◼ Alanine and glutamine are the only two amino

acids released from the muscle in large amounts

◼ Glutamine becomes a major source of energy for

intestinal cells and cells of immune system

◼ Branched-chain α-keto acids become major

sources of glucose and ketone bodies
 Insulin Glucagon
Stimulus Hyperglycemia, Hypoglycemia,
amino acids amino acids,
epinephrine

Effect on memb. Activates Activates

tyrosine kinase adenylate
Receptor cyclase
activity of
insulin receptor

Increases Inhibits
Effect on CHO glycolysis; glycolysis
metabolism stimulates PFK

Stimulates Inhibits
glycogenesis glycogenesis
 References

1. Harper’s Illustrated Biochemistry 30th Edition

by Rodwell, Bender, Botham, Kennelly and Weil
2. Lehninger Principles of Biochemistry 5th Edition
by Nelson and Cox
3. Textbook of Biochemistry With Clinical Correlation
6th Edition by Thomas M. Devlin
4. Lippincott’s Biochemistry 5th Edition
by Champe and Harvey
5. Textbook of Medical Physiology 13th Edition
by Guyton and Hall
» Absorptive state: also called the fed state; the
metabolic state occurring during the first few hours
after ingesting food in which the body is digesting
food and absorbing the nutrients.

» The post—absorptive state lasts until 8 to 12 hours

after your last meal, which is when you enter the
Lette ee
o
® It typically takes 12 hours after your last meal to fully
enter the fasted state.

» When you're in the fasted state your body can burn

fat that has been inaccessible during the fed state.
* Most enzymes show a
“ Allosteric enzymes show a

Enzymes following Michaelis-

Menten kinetics show
hyperbolic curve
<

= 7
= i
3 aa
= “™. Allosteric enzymes
5 show sigmoid curve

3 ——
o

[Substrate]

Figure 4.6
Effect of substrate concentration on
reaction velocity.
Reversible Inhibitors Bind to Enzyme
at Active Site or Alternate site

Substrate Competitive
'
inhibitor

Substrate
Noncompetitive
inhibitor
 4. Effect of substrate concentration

* Most enzymes show a hyperbolic curve

* Allosteric enzymes show a sigmoidal curve

Enzymes following Michaelis-

Menten kinetics show
hyperbolic curve
<

:
:
oe Allosteric enzymes
show sigmoid curve

[Substrate]
o

Figure 4.6
Effect of substrate concentration on
reaction velocity.
 Enzyme regulation

B. Regulation by covalent modification

Covalently modulated enzymes

* Exist in two forms: active and inactive

* The two forms are interconverted by other enzymes

* Reactions involved:
a) phosphorylation — catalyzed by kinase
b) dephosphorylation — catalyzed by phosphatase

* Rx involves —OH group of serine, threonine & tyrosine

Activation/inactivation of glycogen
phosphorylase

Phosphorylase a (active)

) kinase Phosphatase

Beste eee a rt tS ee
Glycogenolysis
 3. Effect of enzyme concentration

The rate of the reaction is directly proportional to the

enzyme concentration at all substrate concentrations.

Amount of enzyme
 Enzyme regulation

3. Induction and repression of enzyme synthesis

Enzymes involved are those needed under selected

physiologic conditions.
Ex. Elevated blood glucose

Increased insulin secretion |

+ Decreased synthesis
Increased synthesis of key of gluconeogenic
enzymes of glycolysis enzyme (PEPCK)
> The basic strategy of metabolism is simple: form ATP,
reducing power, and building blocks for
biosyntheses.
o
» This complex network of reactions is controlled by:

1. allosteric interactions and reversible covalent

modifications of enzymes
2. changes in their amounts
3. compartmentation
4. interactions between metabolically distinct
organs.
» The enzyme catalyzing the committed step ina
pathway is usually the most important control
1 (= O

>» Opposing pathways such as gluconeogenesis

and glycolysis are reciprocally regulated so that
one pathway is usually quiescent when the other
is highly active.
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Glucose Transporter

» Glucose transporter (GLUT) is a facilitative

transport protein involved in glucose translocation
Ted camsom cy Mesa Myc11

> Glucose transporters accomplish the movement of

glucose from the extracellular space (bloodstream)
into cells.

» GLUTs have acquired distinct physiologic and

biochemical properties that allow them to serve
specific functions in the tissues in which they are
expressed.
 Transporter Tissues* Typeof Notes Sensitive to
transport insulin?
SGLT Renal tubules, Secondary Responsible for the absorption (intestine) No
intestinal epithelia active and reabsorption (renal tubule cells) of
(apical membrane) transport glucose.
Pancreatic betacells, Facilitated Pancreatic beta cells: important for gauging
hepatocytes diffusion blood glucose levels in humans.

Hepatocytes: bi-direction transport of

glucose when influenced by hormones, such
as thyroid hormone.
Pancreatic beta-cells, Facilitated +Hepatocytes: important for the bi-
hepatocytes, diffusion directional transport of glucose with regards
intestinal epithelium, to hepatic glucose metabolism.
Kidney
CNS Facilitated Very high affinity for glucose.
diffusion
Skeletal muscle, Facilitated Expression regulated by insulin.
cardiac muscle, diffusion
adipose tissue
*Most transporters are found in a variety of tissues, but are expressed in higher
concentrations in specific cell-types.
 Adipose tissue in Fat Metabolism

«s 3. Decreased lipolysis due to inactivation of

hormone-sensitive lipase
2
- Lipolysis is the breakdown of stored
triglycerides in the adipose tissue releasing
glycerol and free fatty acids.

- Insulin favors the dephosphorylated,

inactive state of lipase
> Muscle tissue is a significant consumer of nutrients
and oxygen.

» During intense exercise muscles alone account for

60 — 90% of total oxygen consumption.

> Muscle cells are also able to function effectively for a

period of time without oxygen supplies —
anaerobically (anaerobic glycolysis)

>» Increased lactic acid production 2

> One of the variables that affect your resting
metabolic rate is the amount of lean muscle you
Eee
oO
» At any given weight, the more muscle on your body,
and the less fat, the higher your metabolic rate.

> That's because muscle uses a lot more energy than

fat while at rest.
>» The metabolic patterns of the brain, muscle,
adipose tissue, kidney, and liver are very
elhatse-ala

>» Glucose is essentially the sole fuel for the

brain in a well-fed person.

> During starvation, ketone bodies (acetoacetate

and 3-hydroxybutyrate) become the
predominant fuel of the brain. ~
> The adaptation to prolonged starvation is
primarily a change in the pattern of ketone use.

> The brain gives up its addiction to glucose,

and survives on a diet of ketones.

> Atthis stage, nothing else uses glucose- every

tissue in the body either burns fatty acids,
burns ketones, or uses the Cori cycle.
Fuel Choice During Starvation.

>» The plasma levels of fatty acids and ketone bodies

increase in starvation, whereas that of glucose
decreases.
2)
> The liver obtains energy for its own needs by
oxidizing fatty acids released from adipose tissue.

» Hormone-sensitive lipase is the key enzyme

involved in lipolysis, the breakdown of triglycerides
to free fatty acids and glycerol.

>» It is a covalently modulated regulatory enzyme that

is activated through phosphorylation catalyzed by a
cAMP-dependent protein kinase A.
 a

The Role of Adipose Tissue in Starvation

Triacyglycerol is the chief storage

form of energy in our body
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10 20 30
Days of starvation

Figure 26.4
Concentrations of fatty acids and
B-hydroxybutyrate in the blood during
starvation.
> Depending on the amounts and types of nutrients
ingested, the absorptive state can linger for up to 4
hours.
o
>» Insulin from the pancreatic islet 8 cells dominates
during the absorptive state.

» The absorptive state of metabolism lasts for about

four hours, during and after each meal.

> Throughout this state, digested food is converted

into sugar or glucose. ...
> During the postabsorptive state, the
digestive tract is empty and €nergy comes
from the breakdown of the body's reserves.

> During the postabsorptive state, when blood

glucose levels drop the hormones glucagon
and cortisol are released.