Sinteza corpilor cetonici

(cetogeneza)

Principala cale de metabolizare a acetil CoA

includerea n ciclul Krebs (n condiiile n
care scindarea lipidelor i a glucidelor este
echilibrat) - lipidele ard n flacra
glucidelor
n lipsa glucidelor; inaniie, diabet - OA se
utilizeaz pentru generarea Gl.
n lipsa OA, Acetil Co A recurge la formarea
corpilor cetonici: acetoacetatul, -

hidrohibutiratul i acetona

Sinteza lor are loc n ficat, dar se utilizeaz

de esuturile periferice
Au rol energetic (muchiul cardiac, stratul
cortical al rinichilor)

cetogeneza

1.

2.

Utilizarea corpilor
cetonici
Acetoacetatul 2 mol de
acetil CoA, utilizate ulterior
n ciclul Krebs (23 ATP)
A doua cale de activare a
acetoacetatului poate fi:
Acetona:
pn la propandiol (CH3-CHOHCH2OH) , scindat la fragmente
acetil i formil
Transformat n piruvat (prin
hidroxilare dubl)

Cetonemie, cetonurie

Cetonemie- mrirea c% de corpi cetonici n

snge
Cetonurie apariia CC n urin
Diete bogate n lipide, srace n glucide;
inaniie, diabet, dereglri gastrointestinale
la copii sau gravide; glucozurie renal
Eliminarea hidroxibutiratului i
acetoacetatului din organism (fiind anioni
la excreie) conduce la pierderea de
cationi Na- rezult cetoacidoza
Pierderea H2O dehidratarea
organismului

Biosinteza
lipidelor

Obiectivele:

1.
2.
3.

Biosintaza acizilor grai:

saturai cu numr par de atomi de carbon;
nesaturai cu numr par de atomi de carbon;
saturai cu numr impar de atomi de carbon.
Enzimele, coenzimele, reglarea.
Biosinteza TAG: substanele iniiale, enzimele i
coenzimele, reglarea.
Biosinteza fosfogliceridelor: substratele, reaciile
pariale ale I i a II ci;
Biosinteza sfingolipidelor: precursorii, reaciile
principale, enzimele, reglarea.
Metabolismul colesterolului. Biosinteza
colesterolului substratele, etapele, reaciile
pariale ale I etape (pn la acidul mevalonic),
enzimele, coenzimele, reglarea. Cile de utilizare
i eliminare ale colesterolului.

Sinteza AG

Sinteza AG i ncorporarea lor n Tg

constituie mecanismul principal de stocare
a excesului de glucide alimentare (Gl nu se
mai transform n glicogen dar n Tg)
Etapele:
Sinteza de novo cu formarea acidului
palmitic
Elongarea acidului palmitic
Introducerea de legturi duble n AG

Particularitile sintezei
AG
Are loc n citozol

1.
2.

E acid gras sintetaza alctuit din 8

proteine (domenii)- 7 sunt enzime, a 8-a
proteina (purttoare) transportatoare de
acil -ACP.
ACP cuprinde 2 grupe SH:
SH furnizat de un rest de cisteinil: SH-Cis
- SH - fosfopanteteina, ataat prin
legtura fosfat-Ser: SH-Pant
Ca iniiator este acetil CoA (rezultat din
glicoliz), pe cnd sursa major malonil
CoA
rolul reductor i revine NADPH+H

Sinteza de novo cu
formarea acidului palmitic

1.

2.
3.

Etapele:
transferul lui Acetil CoA din
mitocondrii n citozol
Sinteza de malonil CoA
Sinteza acidului palmitic

Transferul lui Acetil CoA din

mitocondrii n citozol

Sinteza de malonil CoA

acetil-CoA + HCO3 + ATP ADP +
Pi + malonil-CoA
E- acetil CoA
Carboxilaza
citrat,
Insulina
palmitoil CoA
Glucagonul

Sinteza acidului palmitic

Sinteza acidului palmitic

Sinteza acidului palmitic

Ciclu de reacii este reluat: butiril+ACP

se condenseaz cu malonil+ACP- formnd
n final C6-acil ACP.
Catena AG crete pn la formarea
palmitil-S-ACP

Reacia sumar:
Acetil-ACP+7 malonil-CoA +14
NADPH+H
Palmitat +7CO2+14NADP + + 8HSCoA+6H2O

deoarece malonil CoA se sintetizeaz

din acetil CoA:
8 acetil-CoA + 14 NADPH +H + + 7
ATP palmitate+ 14 NADP+ +
8HSCoA + 7 ADP + 7 Pi

Elongarea AG

Localizat: reticulul endoplasmatic

AG este activat
La acidul preexistent (palmitil CoA) se ataeaz malonil
CoA

Biosinteza AG nesaturai

Pot fi sintetizai AC mononesaturai. Introducerea unei

duble legturi are loc prin aciunea unei monooxigenaze
(introduce gruparea hidroxil), urmat de deshidratare

Acidul linoleic i linolenic sunt eseniali (exogen)

Acidul linoleic se transform n acidul arahidonic conform
reaciilor

Sinteza TAG

1.

2.

2 ci:
calea monoacilglicerolului: are loc n peretele
intestinal (enterocite)din produi absorbii
(resinteza lipidelor).
calea glicerolfosfatului: n toate esuturile
(activ: esutul adipos i ficat)
AG sunt incorporai n TAG sub form activ de
acilCoA:

R-COOH + ATP + HS-CoA +H2O R-CO~SCoA + AMP +

2 Pi
E- acil Co A sintetaza

1. calea
monoacilglicerolului

TG mpreun cu FL,Col, proteine

sunt incorparate n CM i secretai
mai departe n vasele limfatice.

calea glicerolfosfatului

originea glicerol
fosfatului

n ficat:
n esut adipos, ficat

Sinteza glicerofosfolipidelor

2 c de sintez:
Sinteza de novo - utilizeaz ca
intermediar comun acidul fosfatidic
Calea de rezerv o sintez din
produse formate
Particularitatea biosintezei FL este
participarea precursorilor n forme
active de derivai ai citidin fosfatului
(CDP) ca CDP-colina, CDPetanolamina, CDP-diglicerid.

Sinteza de novo

2. sinteza din produse

formate

Sinteza sfingolipidelor

Se formeaz din palmitoil CoA i Ser

Sfingozina liber se formeaz din
ceramid
Sinteza are loc pe suprafaa
citozolic a membranelor reticulului
endoplasmatic

Sinteza sfingolipidelor

Sinteza Colesterolului

1.
2.

3.

Se sintetizeaz din Acetil-CoA

Necesit 18 moli de Acetil-CoA i 18 de ATP
Principalul organ de metabolizare este ficatul,
dar are loc i n intestin, suprarenale,
tegumente
Are loc n 3 etape:
Sinteza acidului mevalonic
mevalonatul prin mai multe reacii - 3izopentenil pirofosfat. 6 molecule de 3izopentenil pirofosfat scualen
Scualenul se supuine ciclizrii lanosterol -- Col

HO
H2C

CH2

CH2

2 ATP
(2 steps)
2 ADP

CH3

CH2

O
CH2

O
O

5-pyrophosphomevalonate

CH3

ATP
ADP + Pi

CO2

C
H2C

OH

mevalonate

HO
H2C

CH3

CH2

CH2

P
O

O
O

P
O

isopentenyl pyrophosphate

C
H2C

CH3

CH3

CH3
CH2 CH2

isopentenyl
pyrophosphate

2 H3C

O
O

CH

CH2

NADPH

CH2

CH3
CH

CH2

CH2 C

O
CH

CH2

2 farnesyl pyrophosphate

P
O

NADP+ + 2 PPi

NADP+
NADPH

H+

CH3
O

C
H3C

CH

CH2

dimethylallyl
pyrophosphate

P
O

NADP+
NADPH

O2

O
O

O2

squalene

2,3-oxidosqualene

2,3-oxidosqualene

19 steps
HO

squalene

HO

H+

H2O

H2O

HO

HO

lanosterol

lanosterol

cholesterol

lanosterol

REGLAREA I
PATOLOGIA
METABOLISMUL
UI LIPIDIC

Obiectivele

Metabolismul eicosanoizilor. Cile ciclooxigenazic i lipooxigenazic ale

biosintezei lor. Inactivarea.
Metabolismul vitaminelor liposolubile: sursele alimentare, necesitile diurne,
transformrile
Reglarea metabolismului lipidelor la nivelul celulei.
Reglarea neurohormonal a metabolismului lipidelor. Rolul lipotropinelor,
ACTH, hormonilor tiroizi, insulinei, glucagonului, glucocorticoizilor i
catecolaminelor.
Relaiile reciproce dintre metabolismul energetic, glucidic i lipidic.
Dereglrile digestiei i absorbiei lipidelor. Steatoreea pancreatic, hepatic
i intestinal.
Dislipidemiile:
a) hipolipoproteinemiile familiale afeciunea Tangier, - i -lipoproteinemia
familial;
b) hiperlipoproteinemiile primare i familiale;
c) hiperlipoproteinemiile secundare (dobndite) n diabet zaharat,
alcoolism,
afeciuni ale glandelor endocrine.
Cauze, mecanismele dereglrii metabolismului lipidelor, manifestrile
biochimice.
6. Lipidozele tiszlare:
a) ereditare Neimann-Pick, Tay-Sachs, Krabbe, Gaucher, Farber,
leucodistrofia
metacromatic, gangliozidoza GM1;
b) dobndite obezitate, ateroscleroz, alcoolism.
Cauze, mecanismele dereglrii metabolismului lipidelor, manifestrile
biochimice.
7. A-, hipo- i hipervitaminozele A, D, E, K cauze, manifestri metabolice.
8. Rolul eicosanoizilor n procesele inflamatorii, reaciile alergice, dereglrile
fluiditii sanguine.

Metabolismul
eicosanoizilor

O
H3C

The input to fatty acid

synthesis is acetylCoA, which is
carboxylated to
malonyl-CoA.

SCoA

acetyl-CoA
O

OOC

CH2

SCoA

malonyl-CoA

ATP-dependent carboxylation provides energy

input.
The CO2 is lost later during condensation with
the growing fatty acid.
The spontaneous decarboxylation drives the
condensation reaction.

Enzyme-biotin
HCO3 + ATP

ADP + Pi
Enzyme-biotin-CO 2
O
ll

CH3-C-SCoA
acetyl-CoA

2
Enzyme-biotin
O

ll

O2C-CH2-C-SCoA
malonyl-CoA

HCO3 + ATP + acetyl-CoA ADP + Pi +

malonyl-CoA

O
O

C
C

NH

CH CH
H2C

CH
S

Carboxybiotin

O
(CH2)4 C

NH

(CH2)4 CH

lysine NH
residue

Biotin is linked to the enzyme by an amide bond

between the terminal carboxyl of the biotin side
chain and the
-amino group of a lysine
residue.
The combined biotin and lysine side chains act as a
long flexible arm that allows the biotin ring to
translocate between the 2 active sites.

Acetyl-CoA Carboxylase, which converts acetylCoA to malonyl-CoA, is the committed step of the
fatty acid synthesis pathway.
The mammalian enzyme is regulated, by
phosphorylation

allosteric control by local metabolites.

Conformational changes associated with

regulation:
In the active conformation, Acetyl-CoA
Carboxylase associates to form multimeric
filamentous complexes.
Transition to the inactive conformation is
associated with dissociation to yield the
monomeric form of the enzyme (protomer).

Phosphorylated protomer of
Acetyl-CoA Carboxylase (inactive)
Citrate

AMP-Activated
Kinase catalyzes
phosphorylation
of Acetyl-CoA
Carboxylase,
causing
inhibition.

Dephosphorylated,
e.g., by insulinactivated Protein
Phosphatase

Palmitoyl-CoA
Phosphorylated, e.g., via
AMP-activated Kinase
when cellular stress or
exercise depletes ATP.

Dephosphorylated Polymer of
Acetyl-CoA Carboxylase (active)
Regulation of Acetyl-CoA Carboxylase

The decreased production of malonyl-CoA prevents

energy-utilizing fatty acid synthesis when cellular
energy stores are depleted. (AMP is abundant only
when ATP has been extensively dephosphorylated.)

AMP-Activated Kinase
has a significant role
even in tissues (e.g.,
cardiac muscle) that do
not significantly
synthesize fatty acids.
In such tissues malonylCoA, produced via one
isoform of Acetyl-CoA
Carboxylase, functions
mainly as an inhibitor
of fatty acid oxidation.

O
H3C

acetyl-CoA
ATP +

HCO3

ADP + Pi

SCoA

Acetyl-CoA
Carboxylase
(inhibited by
AMP-Activated
Kinase)

OOC

CH2

SCoA

malonyl-CoA

When AMP is high (ATP low), malonyl-CoA production is

diminished, releasing fatty acid oxidation from
inhibition. This will lead to increased ATP production.

O
H3C

SCoA

acetyl-CoA
O

OOC

CH2

SCoA

malonyl-CoA

A cAMP cascade, activated by glucagon &

epinephrine when blood glucose is low, may also
result in phosphorylation of Acetyl-CoA Carboxylase
via
cAMP-Dependent Protein Kinase.
With Acetyl-CoA Carboxylase inhibited, acetyl-CoA
remains available for synthesis of ketone bodies, the
alternative metabolic fuel used when blood glucose is
low.

Phosphorylated protomer of
Acetyl-CoA Carboxylase (inactive)
Citrate
Dephosphorylated,
e.g., by insulinactivated Protein
Phosphatase

Palmitoyl-CoA
Phosphorylated, e.g., via
AMP-activated Kinase
when cellular stress or
exercise depletes ATP.

Dephosphorylated Polymer of
Acetyl-CoA Carboxylase (active)
Regulation of Acetyl-CoA Carboxylase

The antagonistic effect of insulin, produced

when blood glucose is high, is attributed to
activation of Protein Phosphatase.

Phosphorylated protomer of
Acetyl-CoA Carboxylase (inactive)
Citrate

Regulation of
Acetyl-CoA
Carboxylase by
local metabolites:

Dephosphorylated,
e.g., by insulinactivated Protein
Phosphatase

Palmitoyl-CoA
Phosphorylated, e.g., via
AMP-activated Kinase
when cellular stress or
exercise depletes ATP.

Dephosphorylated Polymer of
Acetyl-CoA Carboxylase (active)
Regulation of Acetyl-CoA Carboxylase

Palmitoyl-CoA (product of Fatty Acid Synthase)

promotes the inactive conformation, diminishing
production of malonyl-CoA, the precursor of fatty acid
synthesis.
This is an example of feedback inhibition.

Citrate
allosterically
activates AcetylCoA Carboxylase.

[Citrate] is high when there is adequate

acetyl-CoA entering Krebs Cycle.
Excess acetyl-CoA is then converted via
malonyl-CoA to fatty acids for storage.

Fatty acid synthesis from acetyl-CoA & malonylCoA occurs by a series of reactions that are:

in bacteria catalyzed by 6 different enzymes

plus a separate acyl carrier protein (ACP)

in mammals catalyzed by individual domains

of a very large polypeptide that includes an
ACP domain.
Evolution of the mammalian Fatty Acid
Synthase apparently has involved gene fusion.

NADPH serves as electron donor in the two

reactions involving substrate reduction.
The NADPH is produced mainly by the Pentose
Phosphate Pathway.

SH

Coenzyme A

CH2
CH2

-mercaptoethylamine

NH

Fatty Acid
Synthase
prosthetic groups:

the thiol of the sidechain of a cysteine

residue of
Condensing Enzyme
domain.
the thiol of
phosphopantethei
ne, equivalent in
structure to part of
coenzyme A.

CH2
CH2

pantothenate

NH
C

NH2
O

ADP-3'phosphate

HO

H3C

CH3 O

H2C

O
O

CH2

O
H

phosphopantetheine

H
OH

P
O

SH
CH2

Phosphopantethein
e (Pant) is covalently
linked via a phosphate
ester to a serine OH
of the acyl carrier
protein domain of
Fatty Acid Synthase.
The long flexible
arm of
phosphopantetheine
helps its thiol to move
from one active site to
another within the
complex.

phosphopantetheine
of acyl carrier protein

CH2

-mercaptoethylamine

NH
C

CH2
CH2

pantothenate

NH
C

HO

H3C

CH3 O

H2C

NH
O

phosphate

CH2

CH
C

serine
residue
O

As each of the substrates acetyl-CoA & malonyl-CoA

bind to the complex, the initial attacking group is
the oxygen of a serine hydroxyl group of the
Malonyl/acetyl-CoA Transacylase enzyme domain.
Each acetyl or malonyl moiety is transiently in ester
linkage to this serine hydroxyl, before being
transferred into thioester linkage with the
phosphopantetheine thiol of the acyl carrier
protein (ACP) domain.
Acetate is subsequently transferred to a cysteine
thiol of the Condensing Enzyme domain.

acetyl-S-CoA HS-CoA
Pant
SH

Cys
SH

Pant

SH

CO2

malonyl-S-CoA HS-CoA
Cys

S
C
CH3

Pant

Cys

O C

CH2

1 Malonyl/acetyl-CoA-ACP Transacylase COO

2 Malonyl/acetyl-CoA-ACP Transacylase
3 Condensing Enzyme (-Ketoacyl Synthase)

CH3

3
O

Pant

Cys

SH

CH2
C

CH3

The condensation reaction (step 3)

involves decarboxylation of the malonyl
moiety, followed by attack of the resultant
carbanion on the carbonyl carbon of the
acetyl (or acyl) moiety.

NADPH NADP+
Pant

Cys

SH

CH3

Pant

Cys

SH

HC

Pant

Cys

SH

C
CH

CH2

CH2
C

NADPH NADP+

H2O

OH

CH3

HC
CH3

Pant

Cys

SH

CH2
CH2
CH3

4 -Ketoacyl-ACP Reductase
5 -Hydroxyacyl-ACP Dehydratase
6 Enoyl-ACP Reductase
4.
5.
6.

The -ketone is reduced to an alcohol by e

transfer from NADPH.
Dehydration yields a trans double bond.
Reduction by NADPH yields a saturated chain.

Malonyl-S-CoA HS-CoA
Pant

Cys

SH

Pant

Cys

SH

S
C

2
O

Pant

Cys

CH2

CH2

CH2

CH2

CH2

CH2

COO

CH2

CH3

CH3

CH3

7 Condensing Enzyme
2 Malonyl/acetyl-CoA-ACP Transacylase (repeat).

Following transfer of the growing fatty acid

from phosphopantetheine to the Condensing
Enzyme's cysteine sulfhydryl, the cycle begins
again, with another malonyl-CoA.

Product release:
When the fatty acid is 16 carbon atoms long,
a Thioesterase domain catalyzes hydrolysis
of the thioester linking the fatty acid to
phosphopantetheine.
The 16-C saturated fatty acid palmitate is
the final product of the Fatty Acid Synthase
complex.

The primary structure of the mammalian Fatty Acid

Synthase protein is summarized above.
Fatty Acid Synthase in mammals is a homo-dimer.

X-Ray
crystallographic
analysis at 4.5
resolution shows the
dimeric Fatty Acid
Synthase to have an
X-shape, with
domains arranged as
summarized at right.

The solved structure does not resolve the position

of ACP & Thioesterase domains, predicted from
primary structure to be near -Ketoacyl Reductase
(KR) domains of lateral "arms" of the complex.

These domains may

be too flexible to be
resolved.
KR = -Ketoacyl
Reductase; ER = Enoyl
Reductase;
DH = Dehydratase;
KS = -Ketoacyl Synthase
(Condensing Enzyme);
MAT = Malonyl/AcetylCoA Transacylase.

For images see:

website (ETH Zurich)
website (Asturias lab,
Scripps)

article (Maier, Jenni & Ban;

requires subscription to
Science).
Fatty Acid Synthase complex is somewhat
asymmetric.
There is evidence for conformational changes
relating to catalysis.
Protein flexibility may facilitate transfer of ACPattached reaction intermediates among the several
active sites in each half of the complex.

Explore with Chime the structure of

the E. coli -Ketoacyl-ACP Synthase
III, equivalent to the domains of the
mammalian Fatty Acid Synthase that
catalyze the initial acetylation and
condensation reactions.

 -Oxidation & Fatty Acid

Synthesis
Compared
Oxidation Pathway Fatty Acid Synthesis
pathway location

mitochondrial matrix

cytosol

acyl carriers
(thiols)

Coenzyme-A

phosphopantetheine
(ACP) & cysteine

e acceptors/donor

FAD & NAD+

NADPH

-OH intermediate

2-C product/donor

acetyl-CoA

malonyl-CoA
(& acetyl-CoA)

Fatty Acid Synthase is transcriptionally

regulated.
In liver:
Insulin, a hormone produced when blood
glucose is high, stimulates Fatty Acid Synthase
expression.
Thus excess glucose is stored as fat.
Transcription factors that that mediate the
stimulatory effect of insulin include USFs
(upstream stimulatory factors) and SREBP-1.
SREBPs (sterol response element binding
proteins) were first identified for their
regulation of cholesterol synthesis.
Polyunsaturated fatty acids diminish
transcription of the Fatty Acid Synthase gene in
liver cells, by suppressing production of SREBPs.

In fat cells:
Expression of SREBP-1 and of Fatty Acid
Synthase is inhibited by leptin, a hormone
that has a role in regulating food intake and fat
metabolism.
Leptin is produced by fat cells in response to
excess fat storage.
Leptin regulates body weight by decreasing
food intake, increasing energy expenditure,
and inhibiting fatty acid synthesis.

Elongation beyond the 16-C length of the palmitate

product of Fatty Acid Synthase occurs in mitochondria
and endoplasmic reticulum (ER).

Fatty acid elongation within mitochondria involves

the -oxidation pathway running in reverse, but
NADPH serves as electron donor for the final
reduction step.

Polyunsaturated fatty acids esterified to CoA are

substrates for the ER elongation machinery,
which uses malonyl-CoA as donor of 2-carbon units.
The reaction sequence is similar to Fatty Acid
Synthase but individual steps are catalyzed by
separate proteins.
A family of enzymes designated Fatty Acid
Elongases catalyze the initial condensation step for
elongation of saturated or polyunsaturated fatty
acids.

10 9

O
C

OH

oleate 18:1 cis 9

Desaturases introduce double bonds at

specific positions in a fatty acid chain.
Mammalian cells are unable to produce
double bonds at certain locations, e.g., 12.
Thus some polyunsaturated fatty acids are
dietary essentials, e.g., linoleic acid, 18:2
cis 9,12 (18 C atoms long, with cis double
bonds at carbons 9-10 & 12-13).

10 9

O
C

OH

oleate 18:1 cis 9

Formation of a double bond in a fatty acid involves
the following endoplasmic reticulum membrane
proteins in mammalian cells:

NADH-cyt b5 Reductase, a flavoprotein with

FAD as prosthetic group.

Cytochrome b5, which may be a separate protein

or a domain at one end of the desaturase.

Desaturase, with an active site that contains two

iron atoms complexed by histidine residues.

The desaturase catalyzes a mixed function

oxidation reaction.
There is a 4-electron reduction of O2 2 H2O as
a fatty acid is oxidized to form a double bond.
2e pass from NADH to the desaturase via
the FAD-containing reductase & cytochrome
b5, the order of electron transfer being:
NADH FAD cyt b5 desaturase
2e are extracted from the fatty acid as the
double bond is formed.
E.g., the overall reaction for desaturation of
stearate (18:0) to form oleate (18:1 cis 9) is:
stearate + NADH + H+ + O2 oleate + NAD+
+ 2H2O