Glycolysis and

Gluconeogenesis

Glycolysis
What is glycolysis?
sequence

of reactions that converts one

molecule of glucose to two molecules of
pyruvate with the formation of two ATP
molecules
anaerobic

Glycolysis
Why is glucose such a commonly used

fuel?
tends

to exist in ring form, very stable,

doesnt generally glycosylate proteins
formed from formaldehyde under prebiotic
conditions

Glycolysis
What are the possible fates of glucose?

Glycolysis
Whats the difference between a

facultative anaerobe and an obligate

anaerobe?
Can you give an example of habitatdependent anaerobiosis?
What about activity-dependent
anaerobiosis?

Glycolysis
All the intermediates in glycolysis have either

3 or 6 carbon atoms
All of the reactions fall into one of 5
categories

phosphoryl transfer
phosphoryl shift
isomerization
dehydration
aldol cleavage

Glycolysis
Entire reaction sequence may be

divided into three stages

glucose

is trapped and destabilized

six carbon molecule is split into two three
carbon molecules
ATP is generated

Glycolysis Stage 1

glucose converted to glucose-6-PO 4

ATP is needed
catalyzed by hexokinase or glucokinase
induced fit
G01= -4.0 kcal/mole

Glycolysis Stage 1

phosphoglucoisomerase
aldose is converted to ketose
G01=+0.4 kcal/mole

Glycolysis Stage 1

rate limiting enzyme allosteric

inhibited by high ATP, citric acid, long-chain fatty acids
stimulated by ADP or AMP
G01= - 3.4 kcal/mole

Glycolysis

Glycolysis Stage 2

six carbon molecule split into 2- 3 carbon molecules

aldose and ketose
G01=+ 5.73 kcal/mole

Glycolysis Stage 3

At equilibrium most mixture exists as

dihydroxyacetone phosphate
G01=+ 1.83 kcal/mole

Triose Phosphate Isomerase

Glycolysis Stage 3

redox reaction
energy from redox used to form acyl

phosphate
G01= +1.5 kcal/mole

Glycolysis Stage 3
Consists of two coupled processes

Glycolysis Stage 3

formation of ATP substrate level

phosphorylation

Glycolysis Stage 3

phosphoryl shift uses 2,3

bisphosphoglycerate G01= +1.1 kcal/mole

dehydration G01 = +.44 kcal/mole
phosphoryl transfer G01 = -7.5 kcal/mole

Glycolysis

Fate of Pyruvate

Alcoholic Fermentation
Which organisms carry out this process?
yeast
other microorganisms
PDC requires thiamine pyrophosphate as coenzyme
NAD+ is regenerated

Lactic Acid Fermentation

Occurs in muscle cells, microorganisms
Regenerates NAD+

NAD+ and Dehydrogenases

Various dehydrogenases have a similar binding

domain for NAD+ showing their common origin

Rossman fold

Glycolysis
How can fructose be

used for energy?

Glycolysis
To use galactose it must be converted to

glucose-6-PO4

Glycolysis

Glycolysis
What causes lactose intolerance?

Glycolysis
What is galactosemia?
inability

to metabolize galactose
missing galactose 1-phosphate uridyl
transferase
liver disease
development of cataracts
CNS malfunction

Control of Glycolysis
Of what value is glycolysis for cells?
provides energy in form of ATP
provides building blocks for synthetic reactions
Where are most control points found?
enzymes that catalyze irreversible reactions

hexokinase
phosphofructokinase
pyruvate kinase

Phosphofructokinase
Most important control point in

mammalian glycolytic pathway

allosteric

enzyme

activated by AMP and fructose 2,6

bisphosphate
inhibited by high levels of ATP, citrate, fatty
acids

Phosphofructokinase

Hexokinase
Hexokinase is inhibited by its product

glucose-6-PO4
glucose

remains in blood

Glucokinase, an isozyme of hexokinase

is not inhibited by glucose-6-PO4

found

in liver
has lower affinity for glucose

Pyruvate Kinase
Pyruvate kinase exists as isozymes
L form predominates in liver
M form mostly in muscle and brain
PK is an allosteric enzyme

activated by fructose 1,6 bisphosphate

inhibited by ATP, alanine

L form of PK influenced by covalent

modification

inhibited by phosphorylation

Pyruvate Kinase

Glucose Transport
What is the role of glucose transporters

in animal cells?
facilitate

movement of glucose across cell

membrane

What kind of molecule is a transporter

and where is it located?

small

protein embedded in plasma

membrane

Glucose Transport
mammalian glucose transporter

Glucose Transport

Glycolysis and Cancer

Why are rapidly growing tumor cells

dependent upon glycolysis?

insufficient

oxygen supply

What is the function of HIF-1?

hypoxia-inducible transcription factor
stimulates synthesis of many glycolytic
enzymes and GLUT-1 and 3
also stimulates vascular endothelial growth
factor

Gluconeogenesis
What is gluconeogenesis?
synthesis of glucose from non-carbohydrate
precursors
Why is this an important pathway?
What are some of the major precursors?
lactate, amino acids, glycerol
Where does this process occur?
liver, kidney

Gluconeogenesis
If gluconeogenesis involves the conversion of

pyruvate to glucose why is it not simply the

reverse of glycolysis?

glycolysis contains several irreversible reactions

Which reactions in glycolysis are irreversible?

phosphoenolpyruvate to pyruvate
fructose 6-phosphate to fructose 1,6bisphosphate
glucose to glucose 6-phosphate

Gluconeogenesis
What is the first reaction?
O
CH3 CCO2 Pyruvate

+ CO 2 + A TP

biotin
pyruvate
carboxylase
O
CH2 CCO2 - +
CO 2 Oxaloacetate

A DP + P i

Gluconeogenesis
Why is pyruvate carboxylase of special

interest?

structural

properties

contains ATP-grasp domain at N-terminal end

contains biotin-binding domain at C-terminal
end

Gluconeogenesis
What is the role of biotin in this reaction?
prosthetic group lined to -amino group of lysine
residue
carrier of activated carbon dioxide

Gluconeogenesis
Pyruvate

carboxylase is an
allosteric enzyme

activated by acetyl
CoA
needed to form
carboxybiotin

Gluconeogenesis
Carboxylation of

pyruvate occurs in
the mitocondria but
next step in reaction
sequence occurs in
cytosol

Gluconeogenesis
Decarboxylation of oxaloacetate is coupled with
phosphorylation by GTP
enzyme is phosphoenolpyruvate carboxykinase
O
CH2 CCO2 CO 2 Oxaloacetate

+ GTP

OPO 3 2 -

CH2 = CCO2 - + CO 2
Phosphoenol pyruvate

+ GD P

Gluconeogenesis
Which other steps in glycolysis are

irreversible?
conversion

of fructose 1,6-bisphosphate to
fructose 6-phosphate
conversion of glucose 6-phosphate to
glucose

Gluconeogenesis
CH2 OP O3 2 C O
HO
H
H
OH
H
OH
CH2 OP O3 2 -

CH2 OH
fructose-1,6-bisphosphatase
H2 O

Fructose-1,6-bisphosphate

Pi

C O
HO
H
H
OH
H
OH
CH2 OP O3 2 Fructose-6-phosphate

G = -16.7 kJ mol-1
fructose-1,6-bisphosphatase is an allosteric

enzyme, inhibited by AMP and activated by ATP

Gluconeogenesis
Enzyme that catalyzes last reaction not found

in all tissues

liver and kidney cortex

Gluconeogenesis
Is gluconeogenesis an energetically

favorable reaction in the cell?

What drives this reaction?
Are glycolysis and gluconeogenesis

active at the same time?

Regulation of Glycolysis and

Gluconeogenesis
What are some of the factors that

ensure the reciprocal regulation of

these processes?
allosteric

regulators of key enzymes

energy charge
fructose 2,6-bisphosphate
hormones

Regulation of Glycolysis and

Gluconeogenesis

Regulation of Glycolysis and

Gluconeogenesis
fructose 2,6-bisphosphate stimulates

PFK and inhibits fructose 1,6bisphosphase

controlled

by insulin and glucagon and

reflects the nutritional status of the cell

Regulation of Glycolysis and

Gluconeogenesis
How do hormones influence the

enzymes associated with these

processes?
influence

gene expression

change transcription rate

influence degradation of m-RNA

insulin PFK, PK
glucagon PEPCK, fructose 1,6-bisphosphatase

Regulation of Glycolysis and

Gluconeogenesis
What are substrate

cycles and why are

they important?

can amplify
metabolic signals
can generate heat

Regulation of Glycolysis and

Gluconeogenesis
What is the Cori cycle and why is it

important?

Regulation of Glycolysis and

Gluconeogenesis