Chapter

GLUCOhIE,OGENE,SIS

Glycolysis and Gluconeogenesis

Glucose occupies a central role in metabolism, both as a fuel and as precursor of other
biochemicals. The brain and the red blood cells are almost completely dependent on glucose as
an energy source. The liver's capacity to store glycogen is only sufficient to supply with glucose
for about half a day under fasting, intense exercise, low carbohydrate diet or starvation
conditions.(Degradation of glycogen to glucose is the other way of maintaining blood glucose
levets). ln these cases the body's needs must be met by gluconeogenesis (new glucose synthesis)
from noncarbohydrate precursorc. Gluconeogenesis occurs in liver (90%) and, to a smaller extent
in kidney (10%).
The noncarbohydrate precursors that can be converted to glucose include lactate, pyrwate, citric
acid cycle intermediates, and the carbon skeletons of most amino acids (excluding Leu and Lys).
First, all of these substrates must be converted to oxaloacetate, starting substance for
gtuconeogenesis. Gluconeogenesis is highly endergonic. For example, the pathway leading from
phosphoenolpyruvate to glucos+G-phosphate requires 6 molecules of ATP.
Gtuconeogenesis is a pathway consisting of eleven enryme-catalyzed reactions. lt can begin in
the mitochondria or cytoplasm, depending on the substrate being used. Many of the readions are
the reversible steps found in glycolysis. However, gluconeogenesis is not reversible process
contrary to glycolysis. ln glycolisis thrce reactions occur, catalyzed by pyruvate kinase,
phosphofrucfose kinase and hexokinase, which are absent in gluconeogenesis.

(Ihese three reactions are indicated by mark, O, in the next slide..

GLUCONEOGENESIS

pyru vde carborylase

oxaloacetate

Ph osp h

o e no

lopy ruv ate car bory lase

"(*
fructose -116-bis-P

fructose- fr 6 -bis -phosphatase

phosphofructokinase

fructose

-6-

glucose

-6- P

g lu c o s e -6-p h osp h at os e

hexnkinase

@<-

Carboxylation of pyruvate
ln course of gluconeogenesis pyruvate can not be converted to phosphoenolpyruvate
indirectly. GluconCogeneiis begins in the mitochondria with the formation of oxaloacetate through
carboxylation of pyrwate. This reaction also requires one molecule of ATR and is catalyzed by
pyruvate carborylase. This enzyme is stimulated by high levels of acetyl-CoA (produced in F.
oxidation in the liver) and inhibiled by high levels of ADP. ln next step (afrer transfening of
oxaloacetate to cytosol) oxaloace{ate is converted to phosphoenolpyruvate. This way the
irreversible reaction catalyzed by pyruvate kinase is omitted.

.a\'_
v

ATp

H-C_H
!

C:O
I

ADP + Pi

g#::;?,,,

oF-o'

ooc'o'
I

H-C-H
C:O
I

oF-o-

pyruvate

oxaloacetate

fa

\_7

r derived from carboxybiotin

Transport of oxaloacetate from mitochondion to cytosol

Oraloacetate generated in mitochondion must be transferred to cytosol where the enrymes
that convert phospatenolpyruvate to glucose are present.
OxaloacE{ate is tmnsported across milochondrial membrane in twoways.
l.Oxatoacetate must be converted to aspartate to which mitochondrial transport system exist

(malat+aspartate shuttle). This conversion is catalyzed by mitochondrial asparate

aminotansfurase. ln cytosol aspartate is converted back to oxaloacetate by cytosol aspante

aminotansfurase. Next, oxaloacetate is converted phospatenolpyruvate .

2.Oxaloacetate is converted to malate by mitochondrial malate dehydrogenase, which is
oxaloacehte by cytosol malate
cytosol, and converted back
transferred

to

to

dehydrogenase.

flhese two sepante ways are shown on nextslide).

3.Also possible third way of transport of oxaloacetate from mitochondrion to cytosol via citric
acid. lt is mainly used to transport citric acid generated by conversion of lactate.

Oxaloacetate transport from mitochondion to cytosol

Oxaloacetate is transfened from mitochondrion to cytosol by malate. aspartate shuttle. First, in
mitochondrion oxaloacetate must be converted to aspartate or malate (or citrate).
MITOCHONDRION
1 = pyruvate carboxylase

2 = malate dehydrogenaso-

MDH+Ff
acefyl

-SCoA
Eitrate

(mitochondrial)

r?
I
-

3 = malate dehydrogenase-

lt

(cytosol)

*8Sii *8**fr*ffi*]i
uuu lt uuuuilffiilt

ll

I UBU

lt

rcim-l

\'

acetyl SGoAt"/ \

rB*

yt. Frar".e I

*V,
.1f
i
Z

4 = phosphoenolopyruvate

carbo{ykinase
$

asparagate aminotransferEs'

(mitochondrial)
$

asparagate aminotransferase.

(cytosol)

14

co,-v

tcrrosoll

Phosphoenolopyruv

7 = scitrate synthase

= citrate ATP-lyase

DECARBOXYLATION OF OXALOACETATE
oxaloacetate regenerated is decaborylized and phosphatorylated by
energy's donor.
reaction GTP
phoshoenolopyruvate caborykinase.
Phoshoenolopyruvde is formed which (in next step) is converted to fructos+1,6-bis-phosphate in
reversed reaction of glycolysis catalyzed by enrymes taking part in this process.

ln cytosol

is

ln this

H_C-H

H-C-H
il

C=O
I

ooc-o'

phosfroenotopyruvafe
carrboxyklnase

Q:OI

o"c-o'

oxaloacetate

phoshoenolopyruvate

Dephosphorylation of fructose-l,6-bis-phosphate
This reaction is catatyzed by ftucfos*1,&bis phosphatase and allows to omit
irrevercible step of glycolysis (catalyzed by phosphofructokinase.f. Activity of this phosphohse
is regulated by level of cell'energy. High level of AMP inhibits the activity of enryme, when the
high level of ATP together with low level of AMP stimulate gluconeogenesis. This enryme is also
inhibited by fructose.2,6-blb-phoshate (negative allosteric efector) which is regulated by
hormones.

' ./il

H-?-OH

H-C-Or \..-/
C:O

?=o

HO-C-H
H-C-OH
H-C-OH

HO-C-H

H-c:6r\

H-i-o-fil
I-\_/
H

fru ctose-l,6 -bis-p hos ph ate

fru ctose-

1,

6-b i s-ph os ha tas e

H-i\+r/

H-?-o-o
H

fructose6-phosphate

Dephosphorylation of glucose-G-phosphate

Ftydmtysis of glucose-G-phosphate catalyzed by glucose6pfiosphahse allorrus to omit inerenible

step of glycohfsis catalized by hexolcinase or glucokinase. Generate glucose can be transfened to blood and
transported to different tissues .

H. .,O
c

H. ,.O
c

H-C-OH
HO-C-H

H-C-OH
HO-C-H
H-C-OH
H-C-OH
I

H-C-OH +
I

H-C-OH
H-C-OH
I

H-i-o-{il
I-\_/

giluceso-6-phoshatase

glucose

glucose6-phoshate

Gluconeogenesis is very endoergic process. One molecule of glucose is formed from

two molecules of pyruvate . ln this process *3= 6 energy rich bonds are decayed.. ln
gluconeogenesis 4ATP + 2 GTP and2 (NADH + H*) pair is used.
2Pyruvate + 4ATP + 2GTP + 2NADH + 2H*

GIUCOSE

+ 4ADP + zGDP + 2NAD +

6Pi

lntroduction of propionyl residue to gluconeogenesis

FatS acids with uneven number of carbon atoms are converted to n molecules of acetyl
S-GoA and one molecule of propionyl-S-CoA. Propionyl- S-CoA is formed from amino acids
with branched sidechains. e.g. Val and from side chain of cholesterol. Propionyl- S.CoA is
converted to methylmalonyl-S-CoA, which is isomerised to succinyl-S-CoA, which is the
substrate in tricarborylic acid cycle , where via Nastgpnie pzez succinate, fumarate and malate is
converted to oxaloacetate, i. e., a substrate for gluconeogenesis. .

o
C-SoA
il

o
il

C-S-CoA

ArP

C?,

AD

Pi

,-E;*'\\'tsscoA

CoB12

O"C_O-

propionyl-CoA

H_C_H
I

r*rruu^yrc,lB

,-i-,

methylomalonyl-S-CoA

methylmalonyl-S-CoA

o"c-o-

,nuhse
succinyl-S-CoA

COz - from carborybiotin

COB12 - coenzyrne Btz

l0

Regulation of gluconeogenesis

re

tCts

The factors

acetylo--SoA

(enzrymes)

that regulate the

OXALOACETATE

t_

phoshoenolopyruvafe
carboxykinase

particular steps of

I@

gluconeogenesis are
shown in
BLTJE

FRUCTOSE.I,6-biS{P

FRUCTOSEf{

frustose-2,6-b'"@

AMP

fruaos-1,6-bis-

GLUCOSE6

stucos

*i,o- f@

bL-pnospianase

l-_@

11