Metabolism of steroids and other

lipids

CJW

Learning Objectives & Outcomes

This lecture will emphasize on the following topics;
which the students are expected to understand at the
end of this lecture.

1. Biosynthesis of cholesterol.
2. Biosynthesis of steroids, phosphoglycerides &
sphingolipid. Respiratory distress syndrome.
3. Eicosanoids (biosynthesis, roles and applications).

Cholesterol
biosynthesis

Biosynthesis of cholesterol
To synthesize steroids, first we must study the
formation of cholesterol
Cholesterol is synthesized in almost all tissues in
human; predominantly in liver, intestines, adrenal
cortex, reproductive tissues
It is a 27 Carbon molecule
Components: Acetyl-CoA + NADPH + ATP

Way points in cholesterol synthesis

Acetyl-CoA (2C)
HMG-CoA (6C)
Mevalonic acid (6C)
Isopentenyl-pyrophosphate (5C) IPP
Dimethyl allyl pyrophosphate (5C) DAPP
Geranyl pyrophosphate (10C) GPP
Farnesyl pyrophosphate (15C) FPP
Squalene (30C)
Lanosterol (30C)
Cholesterol (27C)

Mevalolic acid / mevalonate

Very similar to fatty acid synthesis,
the building block for cholesterol is
acetyl-CoA

3 Acetyl-CoA (2C) HMG-CoA (6C)

E1: Thiolase (to take away CoA)
E2: HMG-CoA Synthase
Process is driven by the hydrolysis of
CoA high energy thioester bond

Mevalolic acid / mevalonate

Another similarity to FA synthesis;
NADPH is used as a main reductant
E: HMG-CoA reductase
(rate-limiting)
Remove the last CoA
Expression of this enzyme is
inhibited by Cholesterol
Like any feed-back inhibition

Cholesterol synthesis
From mevalonate onwards, to cholesterol
Multi-step process
Points:(1) ATP is used to prime the substrates pyrophosphate
Pyrophosphate compounds are high energy compounds
Its hydrolysis drives pyrophosphatase enzymes
(2) Isopentenyl pyrophosphate (IPP) adds (5C)
(3) NADPH is used to squalene synthesis and cyclization
lanosterol

Steroids biosynthesis
Cholesterol is the precursor to all classes
of steroid hormones
Progesterone
Cortisol (glucocorticoid)
Aldosterone (mineralocorticoid)
Testosterone
Estradiol

Steroid hormones
biosynthesis

Hormone

General principles

Biosynthesis

Transport & Storage

Diversity
Group I

Group II

Action /cascade
Cholesterol biosynthesis

Function
Receptors

Mineralocorticoid
Aldosterone

Glucocorticoid
Types / groups

Regulation

Cortisol

Amino acid modifications

Catecholamines
Dopamine, Norepinephrine
epinephrine

Androgen

Thyronine

Androstenedione

T3 & T4

Testicular
Degradation

Transcription & Translation

Testosterone, DHT

Ovarian
Estradiol, Estrone, Estriol

Other
Calcitriol

Peptide
Insulin
Parathyroid hormone

The steroid hormone family

There are five main steroid hormone family, based on the
number of carbon present as well as the location of its
functional groups

Biosynthesis of
cholesterol
Sources of
cholesterol
1. Plasma, carried by
LDL [main]
2. Synthesized in situ
from acetyl-CoA

Source of cholesterol
The main source of cholesterol for steroid hormone
biosynthesis is transported by LDL and released by
E: Cholesteryl esterase in the cytosol

The cholesterol molecule is transported to the inner

membrane of the mitochondria with the assistance of
Steroidogenic Acute Regulatory (StaR) protein
At the inner membrane of the mitochondria, where
cholesterol is converted to pregnenolone by the
E: cytochrome P450 side chain cleavage enzyme (P450scc)
This is the rate-limiting step for steroidogenesis

Pregnenolone the first product

E: cytochrome P450 side chain cleavage enzyme (P450scc)

Inner membrane of the mitochondria
Rate limiting

Numbering system

Adrenal sterodogenesis
The adrenal gland is one of the main organs that
synthesizes a number of steroid hormones
These include:1. Mineralocorticoid
2. Glucocorticoid
3. Androgen
This is because some of the enzymes required for the
synthesis process is exclusively found in the adrenal glands
(e.g. E: 18-hydroxylase for aldosterone synthesis)

Steroidogenesis: Enzyme and location

(Adrenal)
cytochrome P450 side chain cleavage enzyme
(P450scc)

Inner membrane of mitochondria

3-hydroxysteroid dehydrogenase (3-OHSD)

5,4-isomerase

Smooth endoplasmic reticulum

21-hydroxylase

Smooth endoplasmic reticulum

11 -hydroxylase

Mitochondria

18-hydroxylase

Smooth endoplasmic reticulum

18-hydroxydehydrogenase

Smooth endoplasmic reticulum

17-hydroxylase

Smooth endoplasmic reticulum

17,20-Lyase

Smooth endoplasmic reticulum

Mineralocorticoid
synthesis

Most
potent

Zona glomerulosa

Glucocorticoid
synthesis

Most
potent

Fasciculata & reticularis

zones

Androgen
synthesis
DHEA

Androstenedione

Testosterone

Most
potent

Fasciculata & reticularis

zones

Testicular sterodogenesis
The source of testosterone formation is from
pregnenolone
In the Lydig cells in the interstitial tissue
Follows two pathways:1. Delta 4 (4 pathway) from Androstenedione
2. Delta 5 (5 pathway) the dehydro-epi-androsterone
pathway [common in humans]

Steroidogenesis: Enzyme and location

(Testicular)
cytochrome P450 side chain cleavage enzyme
(P450scc)

Inner membrane of mitochondria

3-hydroxysteroid dehydrogenase (3-OHSD)

5,4-isomerase

Smooth endoplasmic reticulum

17-hydroxylase

Smooth endoplasmic reticulum

17,20-Lyase

Smooth endoplasmic reticulum

17-hydroxysteroid dehydrogenase (17-OHSD)

Smooth endoplasmic reticulum

Testosterone
biosynthesis

Delta 5

Delta 4

(common)
From androgen
synthesis

Reference only

Dihydroxy testosterone (DHT)

Degradation of testosterone is by two pathways;
1st: Mainly oxidation of the 17th C to form 17-ketosteroid
(less active) in many tissues

2nd: At some specific areas: prostate, external genitalia,

some areas of the skin (peripheral); E: 5-reductase to
form DHT (potent)
Finasteride (5-reductase inhibitor)

Dihydroxy testosterone (DHT)

Ovarian steroidogenesis
Follows androgenesis
Produced by peripheral aromatization of androgens
In human males, this accounts for 80% of estrogens found
Aromatization (E: Aromatase) from Androstenedione or
testosterone to form estrone or estradiol, respectively
Estriol is formed by the hydroxylation of the 16th C of
estrone by E: 16-hydroxylase

Biosynthesis of estrogen
Theca cells
Granulosa
cells

Corpus
luteum

Biosynthesis of estradiol
Biosynthesis of estradiol (steroidogenesis); synthesized
from cholesterol (mainly frm: LDL-cholesterol)
Produced in the ovaries; it secretes:Estradiol, progesterone,
testosterone, 17-hydroxyprogesterone
In the ovaries
Pre-ovulation: secretes estradiol
Ovulation: secretes estradiol & progesterone
They are regulated by luteinizing hormone (LH) & follicle
stimulating hormone (FSH)
LH stimulates increase of cAMP increased LDL receptor
increased binding and uptake of LDL-cholesterol

Ovary

1
Cyclophenantane rings

2
3

Precursor to all steroid

hormones

Target for ER+ cancer

therapy

Other types of lipid molecules

Note the sub-groups and over-lapping groups

Respiratory distress syndrome

Affect mostly babies due to the insufficient production
of surfactant in the lungs
Phospholipid (Glycerol + Phosphate + Fatty acids)
Phosphatidyl-choline
Aka. Lecithin

Fn:
most abundant PL in membrane
choline neurotransmission Ach
(acetylcholine)
Surfactant maintains the
dipalmitoyl phosphatidyl choline
elasticity of the alveoli
lung surfactant; prevent alveoli
during breathing
from collapsing (newborn;
respiratory distress)

Respiratory distress syndrome

Usually:
(1) undeveloped lungs
(2) genetic related complications production of
surfactant
Manifestations:
(1) tachypnea (rapid breathing)
(2) tachycardia (irregular heart beat)
(3) chest wall recession
(4) expiratory grunting
(5) cyanosis

Respiratory distress syndrome

References
Harper RK, Granner DK, Mayes, PA, & Rodwell, VW
(2003). Harpers Illustrated Biochemistry (26th Ed).
McGraw-Hill. Chapter 26.

Champe, PC., Harvey, RA & Ferrier, DR. (2011).

Lippincotts Illustrated Review, 5th Ed. LWW. Chapter
18: Cholesterol and steroid metabolism.

Eicosanoids

Learning Objectives & Outcomes

This lecture will emphasize on the following topics;
which the students are expected to understand at the
end of this lecture.

1. Various types of eicosanoids.

2. The general biosynthesis of eicosanoids.
3. Their biological roles & applications

Introduction
Eicosanoids / Prostanoids =
Prostaglandins (PG), Thromboxanes (TX),
Leukotrienes (LT), Prostacyclins (PGI), Lipoxin

They all are derived from 20C polyunsaturated FA

Elicit wide range of responses (physiologic/pathologic)
Produced in a very small amount in ALL TISSUES, short
half-life (rapidly metabolized); Eicosanoids generally act
locally and usually one act antagonistically against the
other

Action usually mediated by G-protein coupled receptors

The precursor (arachidonic acid, 20C) is stored in cell
membrane as phospholipid (arachidonate)

Nomenclature
PGF2 , PGE2, or PGF2
Subscript number refers to the number of double
bonds in the molecule
Sequence: A, B, C. Denotes the functional groups on
the ring structure (cyclo-pentane ring)

Stereochemistry: / designation denotes the

location of hydroxyl group at position 9:
cis to the 11-hydroxyl () or
trans to the 11-hydroxyl ()

Eicosanoids
Prostaglandins (PG)
A,D,E,F
Protacyclins (PGI2)
Thromboxanes (TX)
Leukotrienes (LT)

Subscript denotes the number of

double bonds in it

Fatty acids (FA)

16 to 18 C chains are the most common
Usually they are in even numbers; building blocks,
acetyl-CoA (2C)
Can be unsaturated / poly-unsaturated (usually cis)
naturally occurring

Trans

Cis

Across

On the same side

FA: Nomenclature

18:1;9
1st : Denotes the number of carbon
2nd : Number of double bonds

3rd

Count from
the carboxyl
end

: Locations of the double bonds

1

COOH

18
1. Draw the back-bone and the carboxyl end

FA: Nomenclature (standard)

18:1;9
1st : Denotes the number of carbon
2nd : Number of double bonds
3rd : Locations of the double bonds

1
COOH

18
2. Draw the double bonds

FA: Nomenclature (alternative - omega)

3:18;1
1st : Denotes the location of the first double bond from
the last carbon
2nd : Number of carbons

3rd : Number of the double bonds

1

15

COOH

18
1

3
(omega) ; this is omega-3 (FISH OIL)

Fatty acid synthesis

E: Acetyl-CoA
Carboxylase

E: 3-Ketoacyl-ACP
synthase

E: Malonyl CoA-ACP
tranacylase

E: 3-Ketoacyl-ACP
reductase

1
5

E: Enoyl-ACP
reductase

E: Acetyl-CoA ACP
Transacylase

7
E: Hydrolase/
thioesterase

E: 3-hydroxyacyl-ACP
dehydratase

The seven main steps in fatty

acid synthesis are:1. ACP activation and malonylCoA synthesis
2. Malonyl-transfer (+3C)
3. Condensation
(Decarboxylation) (-1C)
4. Reduction of keto-acyl
(the ketone group)
5. Dehydration
6. Reduction of double bond
(in the growing acyl chain)
7. Hydrolysis

Desaturation
The desaturation process generates various unsaturated
fatty acids
E: desaturase (in the Endoplasmic reticulum)
Requires: NADH and oxygen
Human limitation: (only below 9); 9, 6, 5,4 - desaturases

For references only

Summary: Biosynthesis, Prostaglandin

Stage 1
Release of
arachidonic acid
stored in cell
membrane;
Phospholipase
Stage 2
Prostaglandin
(PGH) synthase
Stage 3
Modifications

Phosphatidylinositol

Synthesis of arachidonic acid (AA)

Synthesized from: Essential FA = Linoleic Acid
From plants
Human X syn; as it involves desaturation of >
9; therefore essential; get it from diet
2 enzymes involved for biosynthesis
a) Elongase (add 2 more C)
b) Desaturase (desaturation; + 2 double
bonds)
Stored in the phospholipid (particularly
phosphatidylinositol)
The release of AA is facilitated by the enzyme
E:phospholipase A2

Synthesis of prostaglandin (PGH2)

Prostaglandin is synthesized from AA by
Prostaglandin endoperoxide synthase
(E:PGH synthase)

It is located at the endoplasmic reticulum

Has 2 catalytic activities:a) Cyclooxygenase (COX) - which cyclicize the
FA; require 2 molecules of O2
b) peroxidase reduces it; requires reduced
glutathion

Isoenzymes of COX
There are two types of COX
COX-1: constitutive in most tissues (always producing)
How asprin involve: gastric tissue, renal homeostasis, platelet
can lead to
aggregation
gastric pain
Inhibitor: Aspirin, Indomethacin, Phenylbutazone
COX-2: non-constitutive (inducible) involve: immune
and inflammatory cells mediates pain, heat,
How
redness, swelling of inflammation
selective
Inhibitor: Aspirin, Indomethacin, Phenylbutazone
inhibitor
prevent
Inhibitor: Selective COX-2 inhibitor (celecoxib;
gastric pain
Celebrex)
Promoter: Cytokines, endotoxin, growth factor,
How cytokines
tumor promotors
induces
inflammation

Inhibition of PGH
synthesis
Corticolsteroids
inhibits the synthesis
of AA; phospholipase
A2
Aspirin inhibits both
COX1/2 affects
stomach, kidney and
impaired clotting
Celebrex is good; COX2 only; but associated
with increased risk of
myocardial infraction

Role of asprin in controling stroke & heart attack

Aspirin inhibits both COX1/2
Anti-thrombogenic effect
Irreversible acetylation of COX-1
(inhibition) in anucleated platelets
Inhibit the production of TXA2 (which
promotes platelet aggregation)
Therefore, low-dose of aspirin is used
to lower the risk of stroke and heart
attacks by decreasing the formation of
thrombi

Thrombosis (blood clot in blood vessel obstruction)

Their roles

Platelet
aggregation
Vaso-motility
Smooth muscle
contraction
Uterine
contraction
Bone metabolism
Nerve and brain
function

*
*

Prostaglandins
Conversion of AA to PGH and TXA

Oxygenases
Arachidonic acid can be oxygenized by TWO enzymes,
besides the E:cyclooxygenase (COX), it can also be
oxidized by E: lipoxygenase
Arachidonic
acid
E:5-lipoxygenase

E:cyclooxygenase
E:peroxidase

5-HPETE
Prostaglandins
Leukotrienes
E:15-lipoxygenase
Lipoxin

E:Prostacyclin
synthase
Prostacyclin

E:Thromboxane
synthase
Thromboxane

Leukotrienes
Besides: AA PGH2 Thromboxanes, Prostacyclins
AA Leukotrienes
They are: Linear hydro-peroxy acids
Eg. In neutrophils (E: 5-lipooxygenase)
AA 5-hydroxy-6,8,11,14 eicosatetraenoic acid
(5-HPETE) series of leukotrienes (A,B,C,D,E)
LTA, LTB, LTC, LTD, LTE
Fn: depends on tissue; generally mediator for allergic
response and inflammation
Leukotrienes inhibitors & receptor antagonist: used in asthma
treatment
There are two other lipoxygenase(s): 12-Lipoxygenase and 15lipoxygenase each inserts the O2 molecule into different
positions

Leukotrienes

Roles of Leukotrienes
Contraction of smooth muscles
Bronchoconstriction
Vasoconstriction
Vascular permeability
Cell adhesion of white blood cells
(counter-infection)

Lipoxins
Derived from
E: 5- & 15-lipoxygenase on AA
Fn: induces chemotaxis and
stimulate superoxide anion
production in leukocytes

Prostanoid therapeutic uses

PGE2
Prostin E2 (oral tablet or suppository)
Induces contraction of the uterine muscles
12-20th week of gestation: elective abortion
Full term: induction of labour
Other similar:
Carboprost (PGF2),
Dinoprost (PGF2)

Prostanoid therapeutic uses

PGE1
Alprostadil (vasodialator)
To treat erectile dysfunction; injected to the corpora
cavernosa

Leukotriene antagonist therapeutic uses

Because leukotrienes are involved in asthma and general
allergies, LT antagonists has therapeutic effect
Eg:
Zileuton (5-lipoxygenase inhibitor)
Montelukast & Zafirlukast
(leukotriene receptor antagonist)

Sources of arachidonic acid / arachidonate

DGLA (20:3 -6)

Evening primrose
oil (EPO)

EPA (20:5 -3)

Fish oil

LTA4
LTB4
LTC4

The importance of diet in

regulating the production of certain
types of prostaglandins

Sources of arachidonic acid / arachidonate

EPA (20:5 -3) & DGLA (20:3 -6) synthesizes Group 1
and Group 3 prostaglandins
Arachidonic acid (from linoleic acid) synthesized
prostaglandins are usually involved in inflammatory
processes
Group 1 & 3 prostaglandins are less pro-inflammatory,
inactive or anti-inflammatory
Therefore it is suggested that -3 (fish) & -6 (EPO) oils
intake has anti-inflammatory effect

Summary

Prostanoid

Types

Nomenclature
Synthesis

Phospholipase A2

PGH Synthase
COX
Prostaglandin, PG
Peroxidase

Drugs

Isomerase/Reductase
Aspirin
Celebrex

PGI synthase

Prostacyclins, PGI

TX synthase

Thromboxane, TX

5-Lipoxygenase

Leucotriene, LT

Seretide
Omega-3 & -6
Others

References
Harper RK, Granner DK, Mayes, PA, & Rodwell, VW
(2003). Harpers Illustrated Biochemistry (26th Ed).
McGraw-Hill. Chapter 23 Metabolism of unsaturated
fatty acids & eicosanoids.
Champe, PC., Harvey, RA & Ferrier, DR. (2008).
Lippincotts Illustrated Review, 4th Ed. LWW. Chapter 17
Complex lipid metabolism.