Oxidation of fatty acid

1
Cont.…
Mobilization of stored fats

• Fatty acids stored in adipose tissue (TAG) coated with

perilipins serve as the body’s major fuel storage reserve

• The mobilization of stored fat requires the hydrolytic

release of fatty acids and glycerol from TAG

• Hormone sensitive lipase (HSL) removes FA from C1

2
Mobilization of triacylglycerols stored in adipose tissue 4
Cont.…
• Hormones that activate HSL
• Catecholamine's
• Glucagon
• Growth hormone
• Glucocorticoids

• Binding hormones to receptors on adipocytes membrane, activate

Adenyl cyclase to produces cAMP which activate Protein kinase (PK)

• A cascade mechanism release FA & glycerol from TAG molecule

5
Cont.…

Regulation of lipolysis
• Insulin decreases lipolysis, but increase esterification of fatty
acids
• By :
❖ Inhibiting adenyl cyclase →→ decreases levels of cAMP

❖ Enhances the uptake of glucose into adipose cells

o α-Glycerol-3-P

6
Cont.…
• When lipolysis is turned on, FA synthesis is turned off

• B/c acetyl CoA carboxylase is inhibited by hormone

directed phosphorylation

• cAMP-mediated cascade is activated as in starvation or

diabetes

• High plasma levels of insulin and glucose in a well fed

state:

• HSL is dephosphorylated (inactive), so lipolysis is turned

off,and fatty acid synthesis and lipogenesis is turned on 7
Summary

8
Cont.…
Fates of glycerol
• Glycerol cannot be utilized by adipocyte due to lacks of
glycerol kinase

• Glycerol is used by the liver to produce glycerol-3-phosphate

❖ glycolysis
❖ gluconeogenesis
❖ synthesis of TAGs
Fates of FFAs
• FFAs are transported in blood in association with serum
albumin-FFAs complex
• Most cells can oxidize FAs to obtain energy (ATP)
• Exception: Brain, Nervous tissues, RBC & Adrenal medulla 9
β-Oxidation of fatty acids
• FFAs are oxidized in tissue to produce energy
• Types
❖ α- oxidation
❖ β- oxidation
❖ ω- oxidation
• β- oxidation is the major method by which FA is oxidized
• There are 3 stages:
❖ Activation
❖ Transport
❖ Degradation
10
Cont.…

[Link]
• Fatty acid is activated to fatty acyl CoA by fatty
acyl CoA synthetase (thiokinase) in the cytosol fatty acyl–CoA
synthetase

AMP + PPi
Phosphatase

11
Cont.…

[Link] into the mitochondria

• Fatty acyl CoA must be transported across the
inner mitochondrial membrane

• Carnitine transports the acyl group from the

cytosol into the mitochondrial matrix

• Carnitine shuttle

• A rate-limiting transport process

12
Role of carnitine in transport of acyl groups
CAT = Carnitine acyl transferase 13
Cont.…
Regulation of CAT I
• CAT-I is related to the outer mitochondrial membrane

• CAT-I reaction is rate limiting

• The enzyme is allosterically inhibited by malonyl CoA

• Malonyl CoA concentration could be high during fatty

acid synthesis

• Inhibition of CAT-I by Malonyl CoA prevents

simultaneous synthesis & degradation of fatty acids
14
Cont.…
Sources of Carnitine

• Diet: meat

• Synthesis in liver & kidney from lysine & methionine

• Skeletal muscle : 97%

• Skeletal & cardiac muscles are entirely depend on

carnitine produced by hepatocytes or taken in diet

15
Cont.…
Deficiency of Carnitine
• Primary cause
❖CAT-II : mainly affects skeletal & cardiac muscle
: inability to use LCFAs

❖CAT-I : affects the liver

: liver can not uses LCFAS to synthesize
glucose during fasting
: fasting hypoglycemia

• Secondary : liver disease, malnutrition, physiologica

change, hemodialysis

• Rx??? 16
Cont.…
3. β- oxidation
• “Mitochondrial pathway, in which 2 carbon fragments are successively
removed from the carboxyl end of the fatty acyl CoA”

• Products
❖ FADH2
❖ NADH
❖ Acetyl CoA

17
Cont.…
• There are four individual reactions each
catalyzed by a separate enzyme

• First Oxidation

• Dehydrogenation between carbon 2 & 3 in

a FAD-linked reaction

• Acyl CoA dehydrogenase

• Produce
❖ trans-∆2-Enol CoA
❖ FADH2 18
Cont.…
• Hydration
• Hydration of the double
bond

• Enoyl CoA Hydratase

• L-3-Hydroxyacyl CoA

19
Cont.…

• Second Oxidation

• Dehydrogenation in a NAD-linked
reaction

• 3-hydroxyacyl Co A dehydrogenase

20
Cont.…
• Cleavage

• Thiolytic cleavage of the thioester

• Beta-ketoacyl CoA thiolase

• Shortened acyl CoA by 2 carbon

atoms

• Acetyl CoA

NB: This 4 steps are repeated until the fatty Acyl chain is completely degraded to Acetyl CoA or
propionyl Co A
21
22
Cont.…
Energy produced from palmitic acid (C-16)
• Each round of degradation produces, one
FADH2, one NADH and one acetyl CoA molecule

8 Acetyl CoA →→ 96 ATP

7 FADH2 →→ 14 ATP
7 NADH →→ 21 ATP
= 131 ATPs
• 2 ATPs utilized to activate fatty acid

• Net gain 131–2

= 129 ATPs
What is the total ATP yield in odd number fatty acid oxidation
23
such as pristanic acid (C-19) ?
α-Oxidation of fatty acids
• Fatty acids with a methyl group at the β-carbon
undergo α-oxidation as methyl blocks β-oxidation

• Phytanic acid presents in milk, animal fats &

chlorophyll

• Site: peroxisomes

• Hydroxylation at the α-carbon by phytanoyl α-

hydroxylase removes C-1

• The product, pristanic acid (C-19), is activated to its

CoA derivative & undergoes β-oxidation
Phytanic acid
24
α-Oxidation of
phytanic acid

Refsum’s disease

25
ω-oxidation
• An alternative minor pathway for fatty acid
degradation

• In ER of liver & kidney

• Preferred substrates: C10 & C12 fatty acids

• Introduces “OH” onto the ω-carbon from O2

• “e” doner NADPH

• Enzymes: cytochrome P450 (mixed function oxidases)

26
27
 Regulation of fatty acid oxidation

• Hormones control the supply of fatty

acids in the blood

• CAT I is inhibited by malonyl CoA

(ACC)

• The rate of ATP utilization controls the

rate of the ETC

• Regulates the oxidative enzymes of -

oxidation & the TCA cycle

28
Coordinated regulation of fatty
acid synthesis and breakdown
29
Ketone bodies

30
Ketogenesis
• During starvation & DM, the acetyl- CoA takes the alternate
fate of formation of ketone-bodies

• Excess acetyl CoA derived from fatty acid, pyruvate or

ketogenic amino acids oxidation have been converted to
ketone bodies in liver

• Ketone bodies are water soluble and an alternative fuel for

cells

Acetoacetate β-3-hydroxybutyrate Acetone 31

Cont.…
1. Two molecules of acetyl-CoA are condensed to acetoacetyl-CoA by
thiolase (acetyl-CoA acetyltransferase)

[Link] of the acetoacetyl-CoA with a third acetyl-CoA by HMG-

CoA synthase forms 𝛃-hydroxy-𝛃-methylglutaryl-CoA (HMG-CoA)

[Link]-CoA is degraded to acetoacetate and acetyl-CoA by HMG-CoA

lyase

• HMG-CoA is also a precursor in cholesterol biosynthesis (Cytosol)

• However, HMG-CoA lyase is present only in liver mitochondria and

therefore does not interfere with cholesterol synthesis
32
33
 Regulation of ketogenesis

• Fasting ⟹glucagon ⟹lipolysis ⟹ ketogenesis

• Acetyl Co A ⟹⇊pyruvate dehydrogenase ,⇈pyruvate

carboxylase

• OAA ⟹ gluconeogenesis (not TCA cycle)

⟹ acetyl CoA ⟹
• ⇊NAD+/NADH ⟹ OAA ⟹malate ketogenesis

36
 Ketolysis
• The ketone bodies are formed in the liver; but they are
utilized by extrahepatic tissues

• Heart muscle and renal cortex prefer the ketone bodies

than glucose

• Skeletal muscle and brain utilize the ketone bodies as

alternate sources of energy, if glucose is not available

• Acetoacetate is activated to acetoacetyl CoA by

Thiophorase (3-ketoacyl-CoA transferase) enzyme

37
38
39
 Ketone bodies are overproduced in DM
and during starvation
• Starvation ⇒ Gluconeogenesis depletes citric acid cycle intermediates

• Untreated DM ⇒ low insulin level leads to inefficient glucose uptake from

the blood
• These conditions leads to low malonyl-CoA, and in turn CPTI free of
inhibition I is relieved

• Fatty acids enter mitochondria degraded to acetyl CoA, and accumulated

• This favors the formation of ketone bodies beyond the capacity of

extrahepatic tissues to oxidize them

• The increased blood levels of acetoacetate and β-hydroxybutyrate lower the

blood PH……….. Acidosis………….. ketosis
40
41
5. Cholesterol Synthesis, transport, & excretion

42
Cholesterol Synthesis
• Cholesterol is a sterol synthesized by virtually all tissues in
humans, but largest contribution are made by:
─ Liver
─ Intestine

• Precursor for:

─ Corticosteroids
─ Bile acids
─ Sex hormones
─ Vitamin D
43
Cont.…
Source of cholesterol
• Diet: 300mg/day

• Synthesis within most cells: 700mg/day

• Precursor: acetyl CoA for all 27 carbon

: ATP and NADPH

• Site: cytosol, endoplasmic reticulum

44
Summary of cholesterol biosynthesis

45
Cont.…
Steps in cholesterol biosynthesis

1. Initial condensation reactions produce HMG CoA, which is

reduced to mevalonate→ Committed Step

[Link] units are formed from mevalonate though loss of

CO2

[Link] isoprenoid units are condensed to form Squalene

[Link] is cyclizes to give rise to parent steroid lanosterol

which is converted to cholesterol by a series of changes
46
I : Acetyl CoA to mevalonate

Committed Step

Statin …………..

47
II Mevalonate to DPP

48
(DPP)
III Formation of squalene

NADP PPi

NADPH + H

49
IV Formation of cholesterol

50
 IV Squalene to cholesterol

Squalene
epoxide

51
52
Metabolic fates of cholesterol

53
 Regulation of cholesterol biosynthesis
1. Hepatic synthesis is inhibited by dietary cholesterol

[Link] CoA reductase is inhabited by cholesterol

[Link] & thyroid hormone increase HMG CoA reductase

activity where as glucagon & glucocorticoids decrease

[Link] CoA reductase synthesis is also controlled at

transcription level

54
Mevalonate
Cholesterol
Bile acid ⊖

Regulation of cholesterol formation 55

6. Lipid transport and storage

56
Cholesterol & other lipids are carried on plasma lipoproteins

• Lipids (C,CE,TAG,PL) are transported in the plasma as plasma

lipoproteins, complexes of specific carrier proteins,
apolipoproteins

• Spherical complexes with hydrophobic lipids in the core and

hydrophilic amino acid side chains at the surface

• However, free fatty acids are carried on albumin

57
58
Cont.…
• Each class of lipoprotein has a specific function, determined by its
point of synthesis, lipid composition & apolipoprotein content

59
Cont.…
• Ten distinct apolipoproteins are found in the lipoproteins of human plasma

60
Cont.…

Lipoproteins Apoprotein
• Chylomicrons A,B-48, C,E
• VLDL B-100,C,E
• LDL B-100
• IDL B-100,E
• HDL A,C,E

61
Cont.…
• Apolipoproteins carry out several roles

❖ Structure of the lipoprotein (apo B)

❖ Cofactors (apo C-II for lipoprotein lipase)

❖ Ligands for lipoprotein receptors ( apo B-100 &

apo E for the LDL receptor)

62
 Chylomicron
• Nanscent chylomicron are synthesized from dietary fats in the ER
of enterocytes, small intestine

• TAGs, PL, Cholesteryl easter, apoB-48 and apo-E enter lymphatic

system,& reach circulation

• apo-C-II and E from HDL make matured chylomicron in

circulation

• ApoC-II activates lipoprotein lipase in capillaries (adipose, heart,

skeletal muscle, & lactating mammary tissues) to release of FFA

63
Cont…
• The remnants of chylomicrons (depleted of most of TAG but
containing cholesterol, apoE, and apoB-48) move through the
bloodstream to the liver

• Receptors in the liver bind to the apoE in the chylomicron

remnants and mediate their uptake by endocytosis

• In the liver, the remnants release their cholesterol and are

degraded in lysosomes

64
Lipoproteins and lipid transport

Chylomicron receptor

65
 VLDL
• Excess fatty acids & carbohydrate from diet are converted into
TAGs in the liver and exported as VLDL to muscle & adipose tissue

• VLDL contains apo B-100, apoC-I, apoC-II, apoC-III, and apoE

• ApoC-II activates lipoprotein lipase, which catalyzes the release of

free fatty acids from TAGs in VLDL

• Adipocytes take up these fatty acids, reconvert them to TAGs

66
 LDL
• The loss of TAGs converts some VLDL to VLDL remnants (intermediate-
density lipoprotein, IDL) in the blood

• Further removal of triacylglycerol from IDL produce LDL

• LDL contains cholesterol, cholesteryl esters & apoB-100 which carries

cholesterol to extrahepatic tissues (muscle, adrenal glands & adipose)

• Plasma membrane LDL receptors recognize apoB-100 and mediate

the uptake of cholesterol and cholesteryl esters

• LDL also delivers cholesterol to macrophages which convert them into

foam cells 67
Indigenous pathway

68
Cont.…
• LDL not taken up by peripheral tissues and cells returns to the liver and
is taken up via LDL receptors in the hepatocyte plasma membrane

• Cholesterol that enters hepatocytes by this path may be incorporated

into membranes, converted to bile acids, or reesterified

• VLDL formation & LDL return to the liver is the endogenous pathway
of cholesterol metabolism and transport

• Accumulation of excess intracellular cholesterol is prevented by

reducing the rate of cholesterol synthesis

69
70
Cont.…
• LDL receptors are synthesized in the Golgi complex and transported to the
plasma membrane, bind apoB-100

• The binding of LDL to an LDL receptor initiates endocytosis, which conveys the
LDL and its receptor into the cell within an endosome

• The receptor-containing portions of the endosome membrane bud off and are
returned to the cell surface, to function again in LDL uptake

• The endosome fuses with a lysosome, hydrolyzes the cholesteryl esters, releasing
cholesterol & FA into the cytosol

• The apoB-100 protein is also degraded to amino acids

• Conversion of VLDL to LDL exposes receptor-binding domain of apoB-100, but

not in VLDL 71
Cholesteryl esters enter cells by receptor-mediated endocytosis
mutation

familial
hypercholesterolemia
(FH)

72
Cont.…
• LDLs are rich in cholesterol and cholesterol esters
❖60% →→ liver to be endocytosed
❖40%→→ extrahepatic tissues (adrenocortex, gonads)

Excess LDL uptake by macrophages

Induce an inflammatory response

Initiate the complex cascade of

atherosclerosis
73
HDL carries out reverse cholesterol transport
• High density lipoprotein (HLDL) originates in the liver and small
intestine

• Contains apoA-I , apoC, apoE, lecithin-cholesterol acyl transferase

(LCAT), little cholesterol & no cholesteryl esters

• LCAT catalyzes the formation of cholesteryl esters from lecithin &

cholesterol

• A major function of HDL is to act as a source for the apoC & apoE for
metabolism of chylomicrons and VLDL

• Nascent HDL consists of discoid phospholipid bilayers containing apo A

and free cholesterol
74
Cont…
• The surface PL & free cholesterol are converted
into cholesteryl esters & lysolecithin

• Cholesteryl esters moves into interior of HDL

disk (Matured HDL)

• HDL becomes spherical due to accumulation of

cholesteryl esters

• Nascent HDL can also pick up cholesterol from

cholesterol-rich extrahepatic cells

❖ Macrophages and foam cells formed from

macrophages
• Mature HDL returns to the liver, unloaded
cholesterol via the scavenger receptor (SR-BI) 75
Cont….
• Excess unesterified cholesterol removed from HDL by:
❖ Scavenger receptor (SR-B1)

oBinds HDL via apo A-I & cholesteryl ester is selectively delivered to
the cells (liver)

oMediates the acceptance of cholesterol effluxes from the cells by

HDL (tissue)

oFree apo A-I is released & destroyed

❖ATP-binding cassette transporters A1 (ABCA1) & G1 (ABCG1)

o ABCG1 mediates the transport of cholesterol from cells to HDL via

coupling the hydrolysis of ATP
76
Metabolism of high-density lipoprotein (HDL) in reverse cholesterol transport 77
Cont….
• The HDL circuit is reverse cholesterol transport

• Depleted HDL then dissociates to recirculate in the bloodstream

and extract more lipids

• Cholesteryl esters from HDL is transferred to VLDL,IDL & HDL

by cholesteryl ester transfer protein (CETP) in exchange for TAG

78
Reverse cholesterol transport
• ApoA-I and HDLs pick up excess cholesterol from peripheral cells, with the participation of
ABCA1 and ABCG1 transporters, and return it to the liver

• Genetically defective ABCA1, the failure of reverse cholesterol transport leads to severe and
79
early cardiovascular diseases: Tangier disease and familial HDL deficiency disease