Androgen Synthesis

C17,20 lyase Dual function protein

Lyase activity important for the
adrenals and the gonads
DHEA Major androgen or androgen
precursor produced by the
adrenal cortex
A pro-hormone
Androstenedione 3β-OHSD; ∆5,4 isomerase
Testosterone Most potent androgen
Produced after the reduction
of androstenedione at C17
by the 17ββ-HSD
The Biochemistry of Hormones, AIA, SY 2007-2008
 ANDROGEN production in women
Adrenal gland

25 % 50 % 90 % 100 %

50 %
Testosterone Androstenedione DHEA DHEA-S

25 % 50 % 10 %

Ovary

The Biochemistry of Hormones, AIA, SY 2007-2008

 ANDROGEN production in women
Sources of Ovaries
Female testosterone 0-1 mg/day
Androgen Androstenedione 1-2 mg/day
production DHEA <1mg/day
Adrenals
DHEAS 6-24 mg/day
Androstenedione 1-2 mg/day
testosterone Very little except if
there are testosterone
producing adrenal tumors

The Biochemistry of Hormones, AIA, SY 2007-2008

 Serum Markers of Female Androgen
production
SOURCE MARKER Remarks:
Ovary TESTOSTERONE NV: 20-80 ng/dl
Adrenals ≥ 200 ng/dl: Androgen
producing tumor
Adrenals DHEA-S ≥ 700 ug/dl: Rare
Imaging is more cost
effective
Peripheral 3 α- DIOL - G Not an absolute
conversion measurement. (20%
overlap)
Diagnosis & therapy
not affected by the
test.
The Biochemistry of Hormones, AIA, SY 2007-2008
The Biochemistry of Hormones, AIA, SY 2007-2008
The Biochemistry of Hormones, AIA, SY 2007-2008
The Biochemistry of Hormones, AIA, SY 2007-2008
 Calcitriol 1,25(OH)2-D3 comes from a
Cholesterol Derivative
Sunlight

7 Dehydrocholesterol Previtamin D3 Vitamin D3

Skin Liver 25-OHlase
Other 25 Hydroxycholecalciferol (25(OH)-D3)
metabolites
24-OHlase 1 α-OHlase

24,25(OH)2-D3 Kidney 1,25(OH)2-D3

Calcitriol
1,25(OH)2-D3

The Biochemistry of Hormones, AIA, SY 2007-2008

 Calcitriol 1,25(OH)2-D3 comes from a
Cholesterol Derivative
7 Dehydrocholesterol Vitamin D3 Calcitriol
1,25(OH)2-D3

The Biochemistry of Hormones, AIA, SY 2007-2008

 Calcitriol 1,25(OH)2-D3
Source Food Fish, liver oil, egg yolk
7 Dehydro- Malphigian layer of the epidermis
cholesterol UV-light proteolysis
Extent of Intensity of light Directly related
conversion exposure
Extent of skin Inversely related
pigmentation
Negative Age-related loss of 7
calcium Dehydrocholesterol in the epidermis
balance and
may be related to the negative
ageing
calcium balance associated with old
age

The Biochemistry of Hormones, AIA, SY 2007-2008

 Calcitriol 1,25(OH)2-D3
Transport protein Vitamin D-binding protein
25 hydroxylation First OBLIGATORY STEP: LIVER (kidney,
intestine)
Occurs in the ER

Co-factors Magnesium
NADPH
Molecular oxygen
Cytoplasmic factor ???
Enzymes P450 reductase
(NADPH)
P450 ???

The Biochemistry of Hormones, AIA, SY 2007-2008

 Calcitriol 1,25(OH)2-D3
25(OH)2-D3 Weak agonist
OH-lation at Mitochondria of the renal PCT
position C1 Enzymes Renal ferredoxin
reductase
required for full
Renal ferredoxin
biologic activity
P450
Co-factor Magnesium
NADPH
Molecular oxygen

The Biochemistry of Hormones, AIA, SY 2007-2008

 TYROSINE DERIVATIVES
1 T3, T4

2 Cathecolamines

The Biochemistry of Hormones, AIA, SY 2007-2008

 Catecholamines
Amines 1 Dopamine
2 Norepinephrine
3 Epinephrine
Epinephrine Major product of adrenal
medulla
80% of all catecholamines
Not made in extramedullary
tissue
Nor-epinephrine 80% made in situ

The Biochemistry of Hormones, AIA, SY 2007-2008

 Catecholamines
Steps in 1 Ring hydroxylation
epinephrine 2 Decarboxylation
synthesis 3 Side chain
hydroxylation
4 N-methylation

The Biochemistry of Hormones, AIA, SY 2007-2008

 Ring Hydroxylation
Tyrosine Immediate precursor
Tyrosine OH-lase rate limiting enzyme
Forms Soluble
Particle bound
Function: Oxidoreductase
Co-Factor:
Tetrahydropterin

The Biochemistry of Hormones, AIA, SY 2007-2008

 Decarboxylation
Dopa decarboxylase Co-Factor:
pyridoxal
phosphate
Competitive
inhibitor: α
methyldopa

The Biochemistry of Hormones, AIA, SY 2007-2008

 Side chain hydroxylation
Dopamine β OH-lase
(DBH)
The Biochemistry of Hormones, AIA, SY 2007-2008
Monooxygenase
Ascorbate: electron donor
Copper: active site
Fumarate: modulator
In the particulate
fraction of the
medullary cells
 N-methylation
Phenylethanolamine-N-Methyltransferase

PNMT Soluble
Phenylethanolamine- Occurs in the cytoplasm of
N-Methyltransferase
the adrenal medulla.
Synthesis is induced
by glucocorticoid
hormones that reach
the medulla via the
intra-adrenal portal
system
The Biochemistry of Hormones, AIA, SY 2007-2008
The Biochemistry of Hormones, AIA, SY 2007-2008
 Triiodothyronine (T3)
Tetraiodothyronine / thyroxine (T4)
Diversity of 1 Requires Iodine for bioactivity
hormones 2 Synthesized as part of a larger
molecule thyroglobulin
3 Stored in an intracellular
reservoir (colloid)
4 Peripheral conversion of T4 to
T3

The Biochemistry of Hormones, AIA, SY 2007-2008

 IODIDE METABOLISM
Steps 1
Concentration
2
Oxidation
3
Iodination
4
Coupling

The Biochemistry of Hormones, AIA, SY 2007-2008

 Oxidation Iodination Coupling
I+ Tgb Tgb Tgb
PEROXIDASE

I-

Phagocytosis
O2 and
NADPH NADP+ Tgb
pinocytosis
H+

Lysosomes
THYROID CELL Tgb

Tgb Secondary
lysosome
Tyrosine

Deiodination Hydrolysis
MIT
I-
DIT
DEIODINASE
I-

Concentration T3T4
Na+-K+ATPase Release
Trans-
porter

EXTRACELLULAR SPACE
The Biochemistry of
I-
Hormones, AIA, SY 2007-2008 T3T4
 THYROGLOBULIN:
A large Prohormone
Precursor of T3 T4
Large, iodinated, glycosylated CHO 8-10%
protein, MW 660kDa Iodide 0.2 – 1%
Two large subunits, 5000 aa
115 tyrosine residues, each a MIT, DIT 70%
potential site of iodination Iodothyronyl 30%
residues T3 T4
Provides conformation required for COUPLING and
IODIDE ORGANICIFACION necessary in the formation
of the diaminoacid thyroid hormones

The Biochemistry of Hormones, AIA, SY 2007-2008

 Oxidation
Facts: Thyroid is the only tissue that can oxidize
I- to a higher valence state.
Obligatory step in I- organification and
TH synthesis
Enzyme: THYROPEROXIDASE
Tetrameric protein, MW 60kDa
H2O2: oxidizing agent
Antithyroid drugs Inhibit oxidation
(thiourea drugs)

The Biochemistry of Hormones, AIA, SY 2007-2008

 COUPLING
FACTS DIT
DIT 4 T
DIT
MIT 3 T
Occurs with the thyroglobulin
molecule
Coupling enzyme????
Thyroperoxidase????

The Biochemistry of Hormones, AIA, SY 2007-2008

 Storage and synthesis
FORM STORAGE RELEASE
Steroid Active None
Hormones form
Catecholamines Active Chromaffin Exocytosis
Supply for
form granules several hours
(Adrenal
Medulla)
T3 and T4 Active Tgb Exocytosis
Supply for weeks
form

The Biochemistry of Hormones, AIA, SY 2007-2008

The Biochemistry of Hormones, AIA, SY 2007-2008
The Biochemistry of Hormones, AIA, SY 2007-2008
 Oxidation Iodination Coupling
I+ Tgb Tgb Tgb
PEROXIDASE

I-

Phagocytosis
O2 and
NADPH NADP+ Tgb
pinocytosis
H+

Lysosomes
THYROID CELL Tgb

Tgb Secondary
lysosome
Tyrosine

Deiodination Hydrolysis
MIT
I-
DIT
DEIODINASE
I-

Concentration T3T4
Na+-K+ATPase Release
Trans-
porter

EXTRACELLULAR SPACE
The Biochemistry of
I-
Hormones, AIA, SY 2007-2008 T3T4
 STEROID METABOLISM

The Biochemistry of Hormones, AIA, SY 2007-2008

 Extraglandular STEROID METABOLISM
Interconversion of
androstenedione
and testosterone
and of estrone and
estradiol
Favors
Increases with advancing
age
Takes place in adipose
tissue
Oxidation of testosterone to
androstenedione
Oxidation of estradiol to
estrone
reduces biological activity

The Biochemistry of Hormones, AIA, SY 2007-2008

 BLOOD TRANSPORT OF STEROIDS
HORMONES
TRANSPORT HORMONE
PROTEIN
SHBG Sex Hormone binding Estrogen, Testosterone,
globulin
Dihydrotestosterone
TEBG Testosterone estrogen
binding globulin

TBG Thyroxine binding globulin T3 T4

CBG Corticotropin binding globulin Cortisol (Hydrocortisone),
/ transcortin
Deoxycortisone,
Progesterone
Albumin DHEA, Androstenedione,
Aldosterone
The Biochemistry of Hormones, AIA, SY 2007-2008
 Transport Proteins
Increased 1 Estrogens
2 Liver diseases
3 Hyperthyroidism

Decreased 1 Androgens
2 Advancing age
3 hypothyroidism

The Biochemistry of Hormones, AIA, SY 2007-2008

 BLOOD TRANSPORT OF STEROIDS
HORMONES
HORMONE FREE ALBUMIN- SHBG-
% BOUND % BOUND %
Estrogen 1 30 69
Testosterone 1 30 69
DHEA 4 88 8
Androstenedione 7 85 8
Dihydrotestosterone 1 71 28

The Biochemistry of Hormones, AIA, SY 2007-2008

 APPROXIMATE AFFINITIES OF STEROIDS
FOR SERUM BINDING PROTEINS
HORMONE SHBG (Kd) CBG (Kd)
Dihydrotestosterone 1 >100
Testosterone 2 >100
Estradiol 5 >10
Estrone >10 >100
Progesterone >100 ≅2
Cortisol >100 ≅3
Corticosterone >100 ≅5

The Biochemistry of Hormones, AIA, SY 2007-2008

 RECEPTORS: Functional Domain
Steroid Binds hormone
Hormones Binds to specific DNA region
Activates/Represses gene transcription
High affinity binding to other proteins
Glucocorticoid Hormone binding region - carboxy
Receptors terminal
DNA binding region
Activates gene transcription (2 regions)
Translocation of receptors from cytoplasm
to nucleus (2 regions)
Binds heat shock protein
The Biochemistry of Hormones, AIA, SY 2007-2008
 Receptors

The Biochemistry of Hormones, AIA, SY 2007-2008

 Cellular mechanism of action

The Biochemistry of Hormones, AIA, SY 2007-2008

 Cellular mechanism of action
Speroff L, et al.
Cir-
Clinical Gynecologic cula-
Endocrinology and tion Cytoplasm Nucleus
Infertility, ed 6,
Lippincott Williams &
Wilkins, Baltimore,
1999

Transcription

mRNA

Ribosomes

Translation

Protein
The Biochemistry of Hormones, AIA, SY 2007-2008
 Cellular mechanism of action
Speroff L, et al. Cir-
Clinical Gynecologic cula-
tion Cytoplasm Nucleus
Endocrinology and
Infertility, ed 6,
Lippincott Williams &
Wilkins, Baltimore, G
r
1999 DNA
o th
w
t
h

mRNA

Re Protein
r cep Micro- synthesis
tor Replenish-
somes
ment

The Biochemistry of Hormones, AIA, SY 2007-2008

 Molecular Mechanism of Action
LEYDIG CELL TARGET CELL

T-R

T T T DHT DHT - R

1. Gonadotropin regulation
2. Spermatogenesis
3. Gene regulation
4. Sexual differentiation
5. Sexual maturation at puberty
6. Gene regulation

The Biochemistry of Hormones, AIA, SY 2007-2008

 Androgen Resistance
LEYDIG CELL TARGET CELL

T-R

T T T DHT DHT - R

Enzyme Deficiencies
1. Side-chain cleavage
2. 17α OH lase
3. Converison of C21 to C19
4. Reduction of 17-ketine
5. Oxidation of A ring to ketosteroid

The Biochemistry of Hormones, AIA, SY 2007-2008

 Cellular mechanism of action
Glucocorticoid
Hormone
Thyroid H and retinoic
acid
hsp Heat shock protein -
+ Co-activator proteins

- Co-repressor proteins
-
Cytoplasm TRE TRE TRE

GRE GRE GRE

hsp
Nucleus
hsp

The Biochemistry of Hormones, AIA, SY 2007-2008

 Phosphoenolpyruvate
The carboxykinase
Biochemistry of Hormones, AIA, SY (PEP-CK)
2007-2008 - a key enzyme in gluconeogenesis