Regulation of Metabolism

Introduction
 The numerous from Stryer 5th ed.

metabolic processes
require tight
regulation

 Within a cell or an
organism, thousands of
different or anta- gonistic
metabolic pathways occur
at the same time

 A precise control and

coordina- tion of the
metabolic pathways is
essential
 Main Tasks of Metabolic Regulation

 Initiation or termination of metabolic pathways:

Depending on the amount and kind of substrates
for anabolism or catabolism, certain metabolic
pathways have to be initiated or terminated

 Coordination of metabolic pathways

The rate of different metabolic processes has to
be adapted to the demands of the organism so
that no product accumulates

 Integration of metabolic pathways

The rate of energy producing pathways has to be
adapted to the rate of energy-consuming pathways
 Principles of Metabolic Regulation

 In the cell
 Compartmentation of metabolic pathways
 Control of enzyme synthesis (expression control)
 Modulation of substrate or cofactor concentration
 Regulation of enzyme activity by covalent
modification or allosteric interactions

 In the whole organism

 Specialization of organs and tissues with respect to
certain metabolic pathways
 Global regulation networks (e.g. by hormones)
 Metabolism as a Regulation Circle
 Control quantity is the flux through a certain metabolic pathway which
has to be kept constant
 Metabolism is controlled by enzymes and global sensory systems
(e.g. hormone-producing glands), which measure substrate or
product concentrations
 Signals are intermediary products (effectors) or signal molecules
(e.g. hormones) which adjust affinity, activity or concentration of
enzymes catalyzing the flux through a metabolic pathway

Control

Detector

Signal

Control quantity

Adjustment
 Regulation by Compartmentation of Metabolic
Pathways

 Compartmentation is an
important way of regulation of
metabolic pathways
 Different metabolic pathways
are often localized in certain
compart- ments (organells)
 Some processes such as
gluconeo- genesis and urea
synthesis depend on the
interplay of reactions that
occur in both compartments
 The fates of certain molecules
depend on whether they are in
the cytosol or in mitochondria;
the flow across the inner
membrane is often regulated
from Stryer 5th ed.
 Principles of Enzyme Mediated Regulation
A
 The flux through a metabolic path-
E1
way depends on the activities of
enzymes catalyzing each reaction B BB
 Some reaction steps are at B B B B B
equilibrium in the cell Enzyme-limited reaction
E2
 enzyme activity is high enough to (far from equilibrium)
equilibrate all the substrate
supplied with the product  C
substrate-limited reactions
E3
 Reactions far from equilibrium are
enzyme-limited due to the low activity Substrate-limited reaction
D (at or near equilibrium)
of enzymes catalyzing these reactions
 These reactions are exergonic, E4
practically irreversible reactions
E EE
 Enzymes catalyzing these exergonic E E E E
E
reactions function as regulatory
valves and determine the flux through E5
the whole pathway
F
 Regulation of Enzyme Synthesis

 The cellular concentration of an enzyme

can be regulated by at least 7 processes
 Synthesis of the primary RNA
transcript
 Posttranslational processing of
mRNA
 mRNA degradation
 Protein synthesis (translation)
 Posttranslational modification of
proteins
 Protein degradation
 Protein targeting and transport
 By expression regulation, a
metabolic pathway can be
completely switched off
 Expression regulation is slow,
compared with activity regulation
 Due to high costs of protein
synthesis, regulation of gene
expression is essential to make
optimal use of available energy
from Lehninger 3rd ed.
 Regulation of Enzyme Synthesis: The Lactose (lac)
Operon
from Stryer 5th ed.

• E. coli can use lactose as carbon

source: lactose is cleaved by -
galactosidase into glucose and
galactose

• In the absence of lactose in the medium,

-galactosidase synthesis is
almost completely switched off

• Addition of lactose to the medium

induces
-galactosidase synthesis

from Stryer 5th ed.

 Regulation of Enzyme Synthesis: The lac Operon

• The genes encoding -

galacto- sidase,
permease, and transace-
tylase are organized in an
operon
• Expression of lacZ, lacY
and lacA is inhibited by a
repressor in the absence
of lactose
• Addition of lactose
(inducer) to the medium
inactivates the repressor from Stryer 5th ed.

and induces expression

 Regulation of Enzyme Activity: Allosteric
Regulation

 The activity of allosterically regulated

enzymes can be increased or
decreased by binding of a modulator
 Modulator can be the substrate or
product of the enzyme-catalyzed
reaction itself or a different regulatory
molecule
 Binding of a modulator to the
regulatory subunit of the enzyme
induces a conformational change of
the catalytic subunit
 The conformational change increases
or decreases the affinity of the
enzyme for the substrate
from Lehninger 3rd ed.
 Regulation of Enzyme Activity: Covalent
Modification
 Another important way of modulating

modified from Lehninger 3rd ed.

enzyme activity is covalent modifica-
tion of the enzyme
Enzyme
 Modifying groups include (less active)
phosphoryl, adenylyl, uridylyl,
adenosine diphos- phate ribosyl,
methyl groups as well as disulfide
bonds Phosphatase Kinase
 These groups are covalently linked
to and removed from the enzyme
by separate enzymes
 The modification induces a
change in the enzyme`s
conformation and can increase
or decrease its activity Enzyme
(more active)
 Protein phosphorylation is the
most important covalent
modification
 Glycolysis and Gluconeogenesis Are Controlled by
Allosteric Interactions and Covalent Modifications
Glucose
ATP Pi
 Glycolysis and gluconeogenesis
are coordinated ADP H2O
Glucose-6-
 One pathway is inactive while the phosphate
other one is active
Gluconeo-
 If both pathways were active at Glycolysis genesis
the same time, the net result
would be the useless 1,3-Bisphospho-
hydrolysis of ATP (futile cycle) glycerate
ADP
ADP
 The rates of glycolysis and
gluconeogene- sis are determined by ATP ATP
3-Phospho-
the concentration of glucose resp. glycerate
the precursor molecules of glucose
 In the whole organism, the rates of
glycolysis resp. gluconeogenesis are Phosphoenol- Oxaloacetate
integrated by the hormones pyruvate
glucagon and insulin, secreted by ADP ADP
the pancreas Pyruvate
ATP ATP
 Glycolysis and Gluconeogenesis Are Controlled by
Allosteric Interactions and Covalent Modifications

 The rates of glycolysis and

gluconeogenesis are controlled
at the committed steps of both
pathways
 A high level of AMP
indicates that energy
charge is low and
stimulates glycolysis
 Conversely, a high level of
ATP signals a high energy
charge and promotes
gluconeogenesis, while
glycolysis is switched off

from Stryer 5th ed.

Allosteric Control of Phosphofructokinase (PFK)
 Phosphofructokinase (PFK)
 Is the key enzyme in the control of glycolysis
 Is allosterically inhibited by high levels of ATP and citrate
 Is allosterically activated by AMP and fructose-2,6-
bisphosphate (F- 2,6-BP)
 F-2,6-BP activates phosphofructokinase by increasing its
affinity for its substrate

Citrate AMP

- +
Fructose-6- + ATP PFK Fructose-1,6- + ADP
phosphate - bisphosphate
+
ATP F-2,6-BP
Pyruvate Kinase Is Regulated by Phosphorylation

 Pyruvate kinase catalyzes the final and third practically irreversible step in
glycolysis
 When the blood glucose level is low, glucagon triggers phosphorylation of
pyruvate kinase, which decreases its activity  the rate of glycolysis
decreases

from Stryer 5th ed.

Ketone Body Formation in the Diabetic State
from Lehninger 3rd ed.
 Ketone Body Formation in the Diabetic
State
 A characteristic metabolic change in diabetes is
excessive oxidation of fatty acids
 The incomplete oxidation results in overproduction of
ketone bodies which exceed the capacity of
extrahepatic tissues
 The oxidation of triacylglycerols to form ketone bodies
produces carboxylic acids which acidify the blood
 This can lead to life-threatening ketoacidosis
Ketone Body Accumulation in Diabetic Ketosis
Urinary Excretion Blood Concentration
[mg/24 h] [mg/100 ml]
Normal < 125 <3
Untreated Diabetes 5000 90
Summary of Clinical and Metabolic Effects of Insulin
Starvation
Effects of insulin starvation on:
Effects of insulin starvation on:
Carbon Metabolism Lipid Metabolism Protein Metabolism
Carbon Metabolism Lipid Metabolism Protein Metabolism
Metabolic deviations glucose utilization lipid synthesis protein synthesis
gluconeogenesis lipolysis protein degradation
Metabolic deviations glucose utilization lipid synthesis protein synthesis
glycogenolysis ketogenesis gluconeogenesis
gluconeogenesis lipolysis protein degradation
Clinical-chemical hyperglycemia
glycogenolysis hyperketonemia
ketogenesis aminoaciduria
gluconeogenesis
findings glucosuria ketonuria hyperglycemia
ketoacidosis glucosuria
Physician`s
Clinical-chemical exsiccosis
hyperglycemia smell of acetone
hyperketonemia muscle degradation
aminoaciduria
findings
findings weight loss
glucosuria weight loss
ketonuria weight loss
hyperglycemia
skin infections telangiectasia
ketoacidosis glucosuria

Physician`s exsiccosis smell of acetone muscle degradation

findings weight loss weight loss weight loss
skin infections telangiectasia