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GLYCOLYSIS

Catabolic pathway, 10 reactions


that occur in the cytoplasm, where
the glucose is degraded to
pyruvate and 2ATP are generated.
Glucose + 2ADP + 2Pi 2Pyruvate +
2ATP
HAD+
H+

NADH +

In anaerobic conditions, pyruvate is


reduced to lactate.
In aerobic conditions, pyruvate is
degraded
by
oxidative
decarboxylation in the citric acid
cycle.
In most tissues, glucose undergoes
aerobic degradation. In muscles,
under extensive work, glycolysis

PREPARATIVE
PHASE
First 3 reactions occurring on
the level of hexose molecule,
which is activated by
phosphorylation. Only the cycic
forms are active!

E1: Hexokinase 4 isoenzymes with different tissue distribution,


specificity, affinity, inhibition by the reaction product, activity, relation
to the blood glucose, insulin effect and biological role.
Glucokinase predominating enzyme in the liver, not inhibited by
glucose-6-P, the activity depends on the blood glucose, insulininduced synthesis. Role: to ensure that the glucose will enter the liver
from blood after feeding.
E2: aldolase intramolecular oxidoreduction.
E3: phosphofructokinase most important allosteric enzyme and the
first specific enzyme for gycolysis.

Degradation of hexose molecules to trioses

Oxidative phosphorylation of glyceraldehyde-3-P,


glycerate to 2-P-glycerate and enolase reaction.

isomerisation

of

3-P

The most important stage of glycolysis 2 oxidative phosphorylations at the


substrate level.

Oxidative phosphorylation of glyceraldehyde-3-P


*

* inhibited by iodoacetate and thus inhibits glycolysis.

Isomerisation

of

3-P

glycerate

to

2-P-glycerate

Enolase reaction
*
Mg2+ / Mn2+

* inhibited by fluoride

Conversion of P-enolpyruvate to pyruvate

Glucose + 2ADP + 2Pi 2Lactate + 2ATP + 2H 2O


Go = - 196 kJ/mol
synthesis of 2 P requires 61 kJ/mol.
Go = 2 x 30.5 = 61 kJ/mol

Efficiency
coefficien
t

Under anaerobic conditions, the reoxidation of NADH


through the respiratory chain to oxygen is prevented.
Pyruvate is reduced by the NADH to lactate, the
reaction being catalyzed by lactate dehydrogenase.
NAD+ is regenerated for oxidative phosphorylation of
glyceraldehyde-3-P.

Under aerobic conditions, pyruvate is taken up into


mitochondria and after conversion to acetyl-CoA is
oxidized to CO2 by the citric acid cycle. The reducing
equivalents from the NADH + H+ formed in glycolysis
are taken up into mitochondria for oxidation via one
of the two shuttles.

Glycerophosphate shuttle functions


in the skeletal muscles and brain.

1 - -Ketoglutarate
transporter;
2 - glu/asp
transporter and
the H+ symport
with glu.

Malate shuttle operates in


myocard, liver and adipose tissue.

Works in
muscle.

heart

and

skeletal

allows rapid transport of highenergy


phosphate
from
the
mitochondrial matrix into the
cytosol.

Cka - concerned with large requirements


for ATP, eg, muscular contraction;
CKc maintains equilibrium between
creatine and creatine phosphate and
ATP/ADP;
CKg - coupling glycolysis
phosphate synthesis;

to

creatine

CKm mitochondrial, mediating creatine


phosphate production from ATP formed
in oxidative hosphorylation;

3M ATP are yielded using malate shuttle to transfer 1M


NADH.

In this case, the complete aerobic degradation of 1M glucose


to CO2 and H2O in gycolysis and citric cycle gives totally 38
M ATP.

2M ATP are generated from 1M NADH


mitochondrion by glycerophosphate shuttle.

that

enter

The complete aerobic degradation of 1M glucose to CO2 and


H2O in gycolysis and citric cycle yield 36 M ATP.

Erythrocytes
mitochondria.

lack

ATP

under

is generated only
anaerobic conditions.
The

first ATP-forming step is


bypassed and energy dissipates as
a heat;
no

net yield of ATP from glycolysis.

However, it does serve to provide


2,3-bisphosphoglycerate,
which
binds to hemoglobin, decreasing its
affinity for oxygen and so making
oxygen more readily available to

The major sites of regulation of glycolysis are the reactions


catalyzed by hexokinase (and glucokinase), phosphofructokinase, and pyruvate kinase.
Phosphofructokinase

tetramer, 2 conformational state T


and R that are in equilibrium.

P is a S and allosteric inhibitor of the enzyme. Each subunit


has 2 ATP-binding sites - substrate that binds both forms in
the same manner, and inhibitory that binds ATP
predominantly in T-conformation. The other S fructose 6-P
binds preferentially to the R-form.
When [ATP] raises, it binds to T-conformation, it moves the
equilibrium in that direction. Thus, the affinity to fructose 6-P
decreases. If the [ATP] is low, the rate of glycolysis increases
and boost up the synthesis of ATP.

Gluconeogenesis - all pathways responsible for converting


noncarbohydrate precursors (first converted to oxaloacetate)
to glucose or glycogen.
Substrates:

glucogenic
glycerol, propionate.
Major

amino

acids

and

lactate,

gluconeogenic tissues - liver and kidney.

Biomedical

importance: meets the needs of the body for


glucose when it is insufficient from the diet or glycogen
reserves (critical for nervous system and erythrocytes).
Failure of gluconeogenesisis usually fatal.
Gluconeogenesis involves reactions from glycolysis,
cytric acid cycle and some special reactions.

Three nonequilibrium reactions catalyzed by hexokinase,


phosphofructokinase, and pyruvate kinase prevent simple
reversal of glycolysis for glucose synthesis.
Phosphoenolpyruvate

(cytoplasmic) Pyruvate (cytoplasmic)

E1: mitochondrial pyruvate carboxylase


E2: mitochondrial and
cytoplasmic
phosphoenolpyruvate
E1
carboxykinase
Mitochondrial ATP and
cytoplasmic GTP
E2

metabolic transportation
between cytoplasm and
mitochondria

E1: Pyruvate carboxylase a tetramer , each unit


containing as a prosthetic group biotin, which side chain is
bound to a lys residue of the enzyme.
Biotin ring is in the end of a long hand that moves it along
the two active center.

1.

In the beginning, 2 is activated by biotinylation with ATP.


A carboxy biotinylated enzyme is received.

2.

Carboxy biotinyllated enzyme transfers 2 to pyruvate,


which in the next step gives oxaloactete.
ATP

Biotinylated enzyme

ADP+P

carboxybiotinylated enzyme

PEPK
PEP

ADP
GDP
ATP
GTP

Pyruvate

Oxaloacetate
Pyruvate

ATP

NADH+H+

Pyruvate
carboxylase

MDH

glu
NAD+
GOT

Malate

MDH

Oxaloacetate

glu

NAD+

Malate
-KG

asp

ADP+P

NADH+H+

PEPK
GOT

-KG

asp

PEP

Fructose-1,6-bisphosphate Fructose-6-P
E1: fructose-1,6-bisphosphatase

Glucose-6-P Glucose
E2:glucose-6-phosphatase
It is present in liver and kidney but absent from muscle and
adipose tissue, which, therefore, cannot export glucose
into the bloodstream.

Energy balance
For the synthesis of 1 mole Glucose by 2 moles Pyruvate
are spent 6 macroergic bonds:
2 moles ATP for the carboxylation of pyruvate to
oxaloacetate;
2 moles GTP to convert oxaloactetate in phosphoenol
pyruvate in carboxykinase reaction;
2 moles ATP for the conversion of 3-P-glycerate to 1,3bisphosphoglycerate in the glyceratekinase reaction.

Changes in the availability of substrates are responsible for most


changes in metabolism (directly or indirectly, acting via changes in
hormone secretion. Three mechanisms are responsible for
regulating the activity of enzymes in carbohydrate metabolism:
changes in the rate of enzyme synthesis;
covalent modification by reversible phosphorylation;
allosteric effects.

Fructose-1,6-bisphosphate is the most important regulated enzyme


of the gluconeogenesis. The direction of the reactions to
glycolysis or to gluconeogenesis, depends on the concentration of
fructose-2,6-bisphosphate. It is a powerful allosteric inhibitor of the
fructose-1,6-bisphosphatase
and
an
activator
of
phosphofructokinase.

The concentration of fructose-2,6bisphosphate depends on the rate


of synthesis and degradation
phosphofructokinase-2
and
fructosebisphosphatase-2.
The concentration of fructose-2,6bisphosphate depends on the rate
of synthesis and degradation
phosphofructokinase-2
and
fructose bisphosphatase-2. Those
two different activities are situated
in two different domains of the
same enzyme, which is regulated
by different allosteric effectors and
by reversible phosphorylation of
protein kinase A.

The Penthose Phosphate Pathway


An alternative route for the metabolism of glucose. It does not
generate ATP but has two major functions:

the formation of NADPH for synthesis of fatty acids and steroids


the synthesis of ribose for nucleotide and nucleic acid formation.
Supplies mast tissue, which does not have glyceraldehyde kinase,
with
glyceraldehyde-3-P
needed
for
the
synthesis
of
triacylglycerols.

Fructose and galactose are converted to glucose, mainly in the liver.

It is more complex pathway than glycolysis.

It occurs in cytosol.

Oxidation occurs by NADP, not NAD+ (glycolysis)

Dehydrogenation and
decarboxylation of glucose 6-P
to yield a pentose, ribulose 5-P.

Ribulose 5-phosphate
is converted back to
glucose 6-phosphate.

1. Glucose-6-P dehydrogenase & 6-P-gluconolactone hydrolase; 2. 6P-gluconolactone dehydrogenase; 3. P-pentoseisomerase; 4. Ppentose epimerase; 5. & 7. transketolase (cofactor TPP); 6.

Net
Balance

3M glucose 6-P give rise to


3M CO2 and 3 5-C sugars.

They are rearranged to


regenerate 2M glucose 6-P
and 1M glyceraldehyde 3P, which is a metabolite of
glycolysis.

Oxidative phase:

Gluconolactone
hydrolase

glucose-6-phosphate
dehydrogenase,
NADP-dependent

gluconolactone
hydrolase - hydrolysis
of 6-P-gluconolactone

6-P-gluconate
dehydrogenase
catalyzes the second
oxidative step

Nonoxidative phase

E1

E2

Ribulose 5-phosphate is the substrate for two enzymes:

E1: Ribose 5-P ketoisomerase converts ribulose 5-P to the corresponding


aldo-pentose, ribose 5-P, which is the precursor of nucleic acid synthesis.

E2: Ribulose 5-P-3-epimerase alters the configuration about carbon 3,


forming another ketopentose, xylulose 5-P.

In fast dividing cells more ribose 5-P is synthesized.

Nonoxidative phase

E1

E2
E1

1.

E1: Transketolase - transfers 2C unit of ketose onto the CHO of aldose


sugar. The reaction requires Mg2+ and co-enzyme TPP(vit B1).

2.

E2: Transaldolase allows the transfer of 3C unit of dihydroxyacetone


moiety from the ketose sedo-heptulose 7-P onto the aldose glyceraldehyde 3-P to form ketose fructose 6-P and 4C aldose erythrose4-P.

Fructose 1,6-bisphosphatase - convert glyceraldehyde 3-P to glucose

In order to oxidize glucose completely to CO 2 via the


pentose phosphate pathway, there must be enzymes
present in the tissue to convert glyceraldehyde 3-P to
glucose 6-P.

The pentose phosphate pathway is active in liver, adipose


tissue, adrenal cortex, thyroid, erythrocytes, testis, and
lactating mammary gland.

Ribose can be synthesized in all tissues.

It is not necessary to have a completely functioning pentose


phosphate pathway for a tissue to synthesize ribose
phosphates.

The Pentose Phosphate Pathway &


Glutathione Peroxidase Protect
Erythrocytes Against Hemolysis

Accumulation of H2O2 may decrease the life span of the


erythrocyte by causing oxidative damage to the cell
membrane.

Pentose phosphate pathway provides NADPH for the reduction


of oxidized glutathione catalyzed by glutathione reductase, a
flavoprotein containing FAD. Reduced glutathione removes
H2O2 in a reaction catalyzed by glutathione peroxidase, an
enzyme that contains the selenium analogue of cysteine
(selenocysteine) at the active site.