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THE UNIVERSITY OF ZAMBIA

SCHOOL OF MEDICINE
DEPARTMENT OF PHYSIOLOGICAL SCIENCES
BIOCHEMISTRY

GLUCONEOGENESIS
Mr Mwale Nicholas K.
Email: mwalenicholask@unza.zm/ mwalenicholask@yahoo.com

OBJECTIVES
Gluconeogenesis
Precursors of gluconeogenesis
Regulation of gluconeogenesis

RESOURCES
All images and notes are taken from the
following unless otherwise stated;
Nelson, D. L & Cox, M. M; Lehninger Principles
of Biochemistry Fifth Edition (2008). W.H.
Freeman & Co. Pp 551-558, 583-590.

Introduction
o Sugar- almost a universal energy source

o Some tissues solely exist on glucose


o Major fuel source for brain, renal medulla

(kidney), embryonic tissues, testes, red blood


cells and nervous system.
o Brain alone requires 120g.
o More than half of all glucose is stored as
glycogen in muscles and liver.

Introduction cont.
Blood glucose and glycogen stores are easily
depleted after periods of fasting or vigorous
exercise.
During these periods, organisms (plants, animals
fungi and microorganisms) need to synthesize
glucose from NON CARBOHYDRATE sources.

Gluconeogenesis= new formation of sugar.

GLUCONEOGENESIS
Important gluconeogenic precursors include;
Lactate
Pyruvate
Glycerol
Certain amino acids
Generally three and four carbon compounds
serve as precursors.
Gluconeogenesis in mammals occurs mainly in the
liver (to lesser extent in kidney and intestinal
wall cells).

GLUCONEOGENESIS cont
New glucose produced goes via blood to supply
tissues
Lactate from skeletal muscle goes via blood to
liver.
In liver this get converted back to glucose and in
muscle will be stored as glycogen.
This cycle is called the Cori Cycle.

Cori Cycle
Active muscles use up
glycogen for energy.
Lactate is generated
during anaerobic
respiration.
Lactate
liver
(generates glucose).
Glucose stored as
glycogen in muscle.

Gluconeogenesis cont.
Occurs in opposite direction to glycolysis (but steps
not completely identical).
7 of 10 reactions use the same enzymes but 3
reactions are irreversible (high negative free
energy change) in glycolysis and use unique enzymes
during the gluconeogenetic pathway;
1. Conversion of phosphoenolpyruvate to pyruvate
2. Conversion of Fructose-6-phosphate to
fructose 1,6-phosphate.
3. Conversion of Glucose to Glucose-6-phosphate

Opposing pathways
of glycolysis and
gluconeogenesis

GLYCOLYSIS Vs GLUCONEOGENESIS CONTD.

Gluconeogenesis cont

In the gluconeogenesis pathway, the 3


irreversible reactions of glycolysis require
enzymes and input of energy.
These enzymes ensure that the reactions are
exergonic and thus irreversible.

Both glycolysis and gluconeogenesis occur in the


cytosol and are regulated by the irreversible
enzymatic stages.

Conversion of pyruvate to PEP

Pyruvate
from
cytosol
is
transported
mitochondria.
Pyruvate can be obtained from alanine
transamination.
Alanine

to

by

When pyruvate or alanine are the glucogenic


precursor, the following reactions are predominant;
Pyruvate
oxaloacetate
phosphoenolpyruvate

Enzymes: pyruvate carboxylase and PEP carboxykinase

Conversion of pyruvate to PEP


cont

First regulatory reaction of pyruvate to oxaloacetate


requires Pyruvate carboxylase (with biotin prosthetic
group).
Acetyl CoA produced during fatty acid metabolism is a
positive effector.
Oxaloacetate produced must be
transported to cytosol.
Done by reducing to malate by
mitochondrial malate DHase then
it passes through the malate
transporter and cytosolic malate
dehydrogenase reconverts malate
to oxaloacetate.

Conversion of pyruvate to PEP cont.


Two high energy phosphate based molecules are
required to convert pyruvate to PEP.
Each yields ~50kJ/mol.
During glycolysis 1 PEP to
pyruvate only uses 1 ATP.
Carboxylation of pyruvate serves
to activate the pyruvate molecule
in prep for subsequent reactions.
If lactate is the pyruvate
precursor a slightly different
path is followed.

Conversion of Fructose 1,6 BP to


fructose 6 P

In glycolysis this is the second irreversible


reaction catalyzed by phosphofructokinase-1.
To reverse this during gluconeogenesis, the
Mg dependent fructose 1,6-bisphophatase is
used here.

Hydrolysis of the phosphate group at C1 is


promoted by this enzyme.

Conversion of Glucose-6 P to glucose


This is the reversal of the hexokinase reaction
involving the dephosphorylation of glucose-6phosphate.
This is energetically unfavorable -requires
transfer of Phosphate to ADP forming ATP.
To overcome this the enzyme glucose-6phosphatase catalyzes the HYDROLYSIS of the
phosphate group.
Enzyme found in liver and renal cells- glucose
made transported to brain and muscle via blood.

Overview of gluconeogenesis
Gluconeogenesis is expensive but highly necessary
for maintenance of blood sugar.
Overall energy expenditure is;

Irreversible reactions commit the substrate into


a singular metabolic pathway.
This also serves to regulate the pathway.

Precursors of gluconeogenesis
4,5,6 carbon molecules from the citric acid cycle
can be oxidized to oxaloacetate.

Precursors of gluconeogenesis
cont
Carbon atoms of many amino acids can

be
catabolized to pyruvate or intermediates of the
citric acid cycle (which are then oxidized to
oxaloacetate).

Precursors of gluconeogenesis
cont..
Acetyl CoA from fatty acid catabolism

in
contrast does not serve as a precursor for
gluconeogenesis.
Amino acids, glycerol and other compounds that
can be catabolized into components for
gluconeogenesis are called glucogenic compounds.

Glycolysis and gluconeogenesis are regulated


such
that
when
glycolysis
is
high
gluconeogenesis is low.
Regulation occurs by allosteric and covalent
modification.

REGULATION OF GLUCONEOGENESIS
Both
glycolysis
(sugar
breakdown)
and
gluconeogenesis (making sugar from non carb
sources) occur in cell cytosol.
Means these two processes are regulated in a
coordinated manner so when glycolysis is
favored, gluconeogenesis is hindered.
The three irreversible reactions serve as a
regulatory point.
Other substrates and by products such as ATP,
AMP, reaction intermediates have an effect on
the enzymes.
Hormones also regulate enzyme activity.

Regulation of gluconeogenesis cont..


Phosphofructokinase-1 which catalyzes the
reaction below is inactive when fructose-2,6bisphosphate is absent.
This occurs even at physiological concentrations
of other allosteric effectors such as AMP.
(AMP signals low cell energy, ATP signals the

opposite)

Effects of F-2,6-BP on
glycolysis and gluconeogenesis

OTHER EFFECTORS INVOLVED IN


GLYCOLYSIS AND GLUCONEOGENESIS

Regulation of gluconeogenesis cont..


The phosphorylated sugar, F-2,6-BP (shown
below) is in turn regulated by a bifunctional
protein.
Catalyzes break down of F-2,6-BP
PFK-2
PFK-2

FBPase-2

Catalyzes formation of F-2,6-BP

PFK-2= Phosphofructokinase 2
FBPase = F-2,6 bisphosphatase

Regulation of Gluconeogenesis cont


Activity of this bifunctional protein depends
on whether it is phosphorylated or not.
Phosphorylation of this protein enhances the
activity of FBPase- (breaks down F-2,6-BPinactivates
PFK-1-glycolysis
stops
and
gluconeogenesis is promoted).

Glucagon hormone-signals low glucose levels


Glucagon activates formation of cAMP
cAMP activates cAMP dependent protein kinases
These kinases phosphorylate the bifunctional
protein.

ACTIVATION/INACTIVATION OF ENZYMES BY
MODIFICATION OF RESIDUES

Effects of hormones insulin and glucagon on


glycolysis and gluconeogenesis

Regulation of gluconeogenesis cont..


cAMP
dependent
protein
kinases
also
phosphorylate liver pyruvate kinases (different
isozyme in muscle) caused by glucagon.
This slows use of glucose for energy and diverts
it to brain and other organs.

In muscle,
cAMP.

epinephrine

(adrenalin)

enhances

Kinases enhanced by this activate glycogen


breakdown (producing glucose for fuel).

Regulation of gluconeogenesis cont


Acetyl CoA
allosterically enhances the
activity of pyruvate carboxylase- promoting
conversion of glucogenic substrates to glucose
(gluconeogenesis).
Acetyl CoA also inhibits the enzyme that
converts pyruvate to acetylCoA (pyruvate
dehydrogenase complex) thus slowing down
the electron transport chain.

Overall high cell energy (NADH, ATP etc.)


reduces glycolysis and favors gluconeogenesis.

Gluconeogenesis and glycolysis are reciprocally controlled

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