Integrated Metabolism

Lecture 02
Prepared by Ishrat Jabeen, PhD

1
 Central Themes of Metabolism

2. How does a cell synthesize the building

1. How does a cell extract energy and reducing
blocks of its macromolecules and then the
power from its environment?
macromolecules themselves?

2
Principles of Human Metabolism
 Human take food as source of chemical energy, and convert it to other
+ forms of energy to:
 Maintaining life e.g. vital work from unconscious part of the brain:
 breathing, pumping blood around the vascular system, chewing food and digesting it,
pumping of ions across membranes, synthesis and breakdown of chemical constituents
of cells.
 And also physical work and nurturing offspring.
 Food can be classified as macronutrients and micronutrients.
 Macronutrients are those components present in a typical serving in amounts of grams
rather than milligrams or less e.g. carbohydrate, fat, and protein.
 Micronutrients are vitamins, minerals, and nucleic acids play vital roles in the
metabolism of the macronutrients
 Water is another important component but it is not nutrient.

3
+ Within body macronutrients are broken down to smaller components,
rearranged, stored, released from stores, and further metabolized
 Thermic effect of food: The energy needed by the digestive system to
process food for storage and use. ~10% of the energy value of foods
consumed is needed to meet this demand.
 Carbohydrates and fats have a thermic effect of about 5% whereas protein is about
30%.
 Thus to metabolize 100 Kcal of carbohydrate, 5 Kcal kilojoules is needed, leaving
+ the body with 95 Kcal of useful energy.

4
 Metabolic pathways are also regulated in common ways. The
principles are
1.Fuels are degraded and large molecules are constructed step by step
in a series of linked reactions called metabolic pathways.
2.An energy currency common to all life forms, adenosine triphosphate
(ATP), links energy-releasing pathways with energy-requiring pathways.
3.The oxidation of carbon fuels powers the formation of ATP.
4.Although there are many metabolic pathways, a limited number of
types of reactions and particular intermediates are common to many
pathways.
5.Metabolic pathways are highly regulated.

5
 Biochemical Energy

Organisms are open systems thus energy and matter can be transferred between the system and its
surroundings

Living cells convert organized forms of energy to heat

A living system’s free energy is energy that can do work when temperature and pressure are uniform, as in a
living cell

6
 Energy Currency of the Cell
 Metabolic pathways can be classified as either:
 energy generating (catabolic)…or
 energy utilizing (anabolic)
 The cell also uses two types of energy currency:
(1) Phosphate anhydrides (compounds with high phosphate transfer potential)
 General: hydrolysis of a phosphate ester

7
 ATP is the most commonly used compound with high phosphate transfer
potential
 phosphoenolpyruvate & creatine phosphate have enough energy to synthesize
ATP
 ATP can be used to synthesize glucose 6- phosphate & similar compounds
 GTP is sometimes used in place of ATP
 ATP is called the energy currency of the cell

8
(2) Reducing equivalents (compounds with high
electron transfer potential)
• Foods → NADH and FADH2 → ATP for Biosynthesis
• We eat reducing equivalents in the foods!!
• In summary- catabolic pathways produce NADH, FADH2, &
ATP; anabolic pathways utilize ATP, NADPH, & FADH 2

9
 Metabolic Pathways Contain Many Recurring Motifs

Metabolism is characterized by common motifs. A small number of recurring activated

carriers, such as ATP, NADH, and acetyl CoA, transfer activated groups in many
metabolic pathways.

NADPH, which carries two electrons at a high potential, provides reducing power in the
biosynthesis of cell components from more-oxidized precursors.

Many activated carriers are derived from vitamins, small organic molecules required in the
diets of many higher organisms.

Moreover, key reaction types are used repeatedly in metabolic pathways.

10
Regulation of Metabolism

+ The amounts of some critical enzymes are controlled by regulation of the rate of
synthesis and degradation.
+ In addition, the catalytic activities of many enzymes are regulated by allosteric
interactions (as in feedback inhibition) and by covalent modification.
+ The movement of many substrates into cells and subcellular compartments also is
controlled.
+ The energy charge, which depends on the relative amounts of ATP, ADP, and AMP,
plays a role in metabolic regulation.
+ A high energy charge inhibits ATP-generating (catabolic) pathways, whereas it
stimulates ATP utilizing (anabolic) pathways.
11
Organ Specialization

 Organs may contain more than one type of tissue, and tissues may
contain more than one cell type.
 Kidney is composed of two distinctive tissue types, the cortex and medulla, and
each of these is composed of various types of cell.
 Skeletal muscle and adipose tissue are tissues, but are
arranged in
+ discrete groups (muscles or fat depots)
 Allof one type of tissue in the body behaves in a broadly similar
manner, and it possible to generalize tissue’s metabolic pattern.
 Various organs and tissues are interconnected by blood vessels to
interact metabolically.

12
Liver Metabolism

 Liver is the first organ to get the nutrients which enter the body
from the intestine after a meal, major role in energy storage after a meal-
 storage of carbohydrate and release of glucose are major functions, and also has
an important role in amino acid metabolism.
 most dietary fat bypasses the liver does not have important roles in fat
metabolism.
+ Dietary short- and medium-chain fatty acids reach the liver directly in portal vein.

13
Carbohydrate Metabolism in the Liver

 Fed Conditions:
 Glucose is absorbed from the intestine into the portal vein, and
hepatocytes (periportal cells) exposed to high glucose
concentrations.
 Liver cells have GLUT-2 glucose transporter not responsive to
insulin, and has a high Km for glucose, so it normally operates
well below saturation.

14
Outline of glucose metabolism and its hormonal regulation in the
liver

Dashed arrows in pathways

indicate multiple enzymatic steps.

GLUT2, hepatic glucose

transporter; G 6-P, glucose 6-
phosphate; GK, glucokinase;
G-6-Pase, glucose- 6-
phosphatase; LDH, lactate
dehydrogenase; PDH,
pyruvate dehydrogenase;
Ribose 5-P, ribose 5-
phosphate; TCA cycle,
tricarboxylic acid cycle.

15
 Within hepatocyte, glucose is phosphorylated to form glucose 6-phosphate
(initial step for metabolism) by glucokinase (hexokinase Type IV),
 It differs from the hexokinases found in muscle and other tissues, that
 It has high Km for glucose (12 mmol/l) like GLUT2; not inhibited by glucose 6-
phosphate, at physiological concentrations. (it is regulated by fructose 6-phosphate)
 Like the GLUT2 transporter it has a high capacity (high Vmax) and is unaffected
by
+ insulin in the short term,
 The presence of the high-Km glucose transporter GLUT2 and the high-Km
glucokinase would NOT enable hepatocyte to take up unlimited quantities of
glucose as glucose 6-phosphate accumulate within cell until
phosphorylation ceased (?)
 There is coordinated control of glycogen synthesis and breakdown: when
one process is stimulated, the other is inhibited.

16
Hormonal Regulation of
Glycogen Breakdown
(Glycogenolysis) and
Synthesis (Glycogenesis)
in the Liver

17
 Glucose binds to a specific site on phosphorylase a, causing a
conformational change that makes the enzyme a better substrate for
dephosphorylation by protein phosphatase-1G.
 Thus, in the liver, an increase in the intracellular glucose concentration will
itself lead to inactivation of phosphorylase.
 The pathways are similar in muscle but there are differences in regulation
 Muscle glycogen breakdown is more susceptible to allosteric effects of AMP
(activation) and glucose 6-phosphate (inhibition).
 Liver glycogen breakdown respond more to stimuli from outside the cell (i.e.,
hormones and the glucose concentration).
 Glycogen synthase in muscle is also phosphorylated by PKA. Therefore, in muscle
adrenaline may also act via PKA to inhibit glycogen synthesis.

18
 Glucose 6-phosphate can also be metabolized via glycolysis to pyruvate
in hepatocytes (activated in fed conditions).
 Some pyruvate may be oxidized directly in the tricarboxylic acid cycle, some
released after conversion to lactate.
 Most of the energy required by the liver for its multiple metabolic
purposes is, derived from the oxidation of amino acids and fatty acids
rather than glucose.

19
c

The Pathways of Glycolysis

and Gluconeogenesis and
Their Hormonal Regulation

20
 Pyruvate carboxylase is present within the mitochondrion, but
PEPCK in
+ humans is equally distributed between cytosol and mitochondrion.
 The three major substrates for gluconeogenesis are: Glycerol,
Alanine, Lactate. They arise from tissues outside the liver.
 Thus the gluconeogenesis is controlled by two major factors: rate of
supply of substrate, and hormonal regulation of the enzymes
 Insulin down-regulate gene expression of some enzymes of gluconeogenesis
 Exercise  blood lactate -------------------------------------
Some reconverted to glucose in
 Starvation  blood glycerol (adipose tissue lipolysis)  the liver

21
 In general, glycolysis is favoured under “fed” conditions, gluconeogenesis under
“starved” conditions. (catalyse opposite functions, and conditions)
 There are three major modes of regulation mediated by hormones:
 allosteric,
 covalent (phosphorylation; through adenylyl cyclase/ cAMP), and
 gene expression (long term regulation, hours rather than minutes) affects GK, G- 6-Pase, and
PEPCK activities. It also involves cAMP, via cAMP-sensitive transcription factors.
 The principal features of regulation of the pathways are as follows:
 F 6-P  allosterically inhibit Glucokinase (to limit glycolysis). But Fructose
+ metabolism  F 1-P  relieves the inhibition of glucokinase.
 F 2,6-P2  + PFK and inhibits FBPase; thus, when
 glucagon  PKA  phosphorylates bifunctional enzyme 6-
phosphofructo-2-kinase/fructose-2,6-bisphosphatase (BFE)  catalyzes breakdown of F
2,6-P2  gluconeogenesis
 insulin  protein phosphatase 2A  dephosphorylates BFE 
+ catalyzes formation of F 2,6-P2  glycolysis

22
  glucagon  +PKA  phosphorylates and inhibits PK ; insulin inhibits this
phosphorylation (i.e., maintains the enzyme active).
 glycolytic flux  F 1,6-P2  allosterically + PK
 Other compounds involved in allosteric regulation are:
 ATP/ADP ratio  glycolysis;
 fatty acids oxidation  citrate (also ATP/ADP ratio)  inhibits
PFK  gluconeogenesis.
 In certain “stress” conditions adrenaline and noradrenaline bind to β-
adrenergic receptors may play the equivalent role of glucagon (i.e. c AMP
concentrations)

23
Overnight Fasted Conditions:
 After meal absorption completed, brain, muscle use glucose, and
blood level begin to fall, slightly; and pancreas favors glucagon secretion.
 Anatomical relationship of the liver to pancreas means hepatic metabolism
is directly regulated by the balance between insulin and glucagon.
 Glucose in blood  Glycogen broken down started

Glucose leave hepatocyte via transporter GLUT2,

and can also transport in vesicles from
endoplasmic reticulum (where it is formed) to
plasma membrane.

24
Glucose Paradox Phenomenon

 Suppress gluconeogenesis while substrate supply increases after a meal.

 Experiment with isolated perfused liver showed the highest rates of glycogen synthesis are not
observed when glucose alone is high, but when glucose is supplied together with lactate, a
precursor of gluconeogenesis
 Lactate must first be converted to glucose 6- phosphate by gluconeogenesis
 Relationships between different organs and tissues are not reproduced in laboratory.
 In humans: hepatic glycogen synthesis after a meal comes about by a combination of
 The “direct pathway” (glucose uptake, glucose 6-phosphate formation, glycogen
+ synthesis) and
 The “indirect pathway” (uptake of three-carbon gluconeogenic substrates, particularly lactate,
formation of glucose 6-phosphate and glycogen synthesis).

25
+ The origin of lactate in this situation is not yet clear. Suggestions are: (a) small
intestine itself, during the process of glucose absorption, (b) some hepatocytes
produce lactate while others use it for gluconeogenesis. (c) Other tissues; RBC,
adipose tissue, and muscle.
+ Not clear the importance of any particular tissue in the postprandial period.
 Pathways around pyruvate carboxylase (PC) and (phosphoenolpyruvate
carboxykinase) (PEPCK)
 Pyruvate enter the TCA cycle and follow two ways of metabolism.
1) Pyruvate dehydrogenase (PDH) convert pyruvate to acetyl-CoA and
combine with oxaloacetate to form citrate and leads to complete oxidation
(i.e. complete oxidation of glucose) or citrate can enter cytosol for
lipogenesis
2) PC convert pyruvate to oxaloacetate may leads to the start of
gluconeogenesis.
26
The Ins and Outs of
the TCA Cycle, and
the Enzymes Pyruvate
Carboxylase and
Phosphoenolpyruvate
Carboxykinase.

27
The pentose phosphate pathway and its links with lipogenesis

28
Fat Metabolism in the Liver

Overview of fatty acid

metabolism in the liver.

29
Fatty Acid Oxidation:
 The liver oxidize fatty acids by the mitochondrial β-oxidation pathway
to produce energy for its many metabolic activities.
 An alternative β-oxidation pathway in peroxisomes operates mainly
to shorten very-long-chain fatty acids, contribute 5-30% of the total hepatic fatty
acid oxidation,
 Inhibitors that prevent fatty acid oxidation gluconeogenesis OR
 If fatty acid supply to the liver  gluconeogenesis
 Gluconeogenesis, requires energy and reducing equivalents
(NADH),
appears to be “fuelled” by oxidation of fatty acids.

30
 Detail of the transport
of fatty acids into
mitochondria

ACS, acyl-CoA. synthase; CPT, carnitine-palmitoyl transferase;

BP, binding proteins for fatty acids (FA) and acyl-CoA.

31
The pathway of ketone
body formation from acetyl-
CoA (ketogenesis).

32
 Lipid synthesis

ACC, acetyl-CoA carboxylase;

FAS, Fatty acid synthase;
HMG-CoA, 3-Hydroxy-3-
methylglutaryl- CoA; OAA,
oxaloacetate; PC, pyruvate
carboxylase; PDH, pyruvate
dehydrogenase.

33
 Outline of amino
acid
metabolism in
the liver.

34
 Outline of the supply of
nitrogen atoms to the
urea cycle.

35