Biochemistry of

Hormones
Part I
Rita Maria Chidiac, M.D.
Chief, Division of Endocrinology
St Georges Hospital-University of
Balamand

„ Historical Background
„ Overview
+ Classes of Hormones
+ Classes of Receptors
+ Neuroendocrine Regulation
+ Hormones & the Hormonal cascade system
„ Synthesis of Polypeptide & Amino Acid-Derived
Hormones
„ Cyclic Hormonal Systems
„ Membrane Hormone Receptors
+ Internalization of receptors
„ Intracellular Hormone Cascade: Protein Kinases
„ Steroid Hormones
+ Biosynthesis/ Regulation of Steroid Hormones
+ Metabolism & Transport of Steroid Hormones
+ Steroid Hormone Receptors
 Discovery & Purification of Hormones
„Historical Background: Insulin discovery
+ 1889 Oskar Minkowsky
+ 1921 Banting & Best

Insulin purification

Nobel prize

1920’s – 1st commercially available insulin

Timeline – 1920s

1922
Charles Best and Frederick
Banting of the University of
Toronto to isolate and purify
insulin, leading to production of
the world’s first insulin therapy.
Insulin & its Metabolic Effects
Isolated from the pancreas in 1922 by Banting
and Best

The endocrine system of the pancreas. In addition to the exocrine cells secreting digestive enzymes, the
pancreas contains endocrine tissue, islets of Langerhans (α, β, & δ cells secreting a specific
polypeptide hormone).
Chemical Structure & Synthesis of Hormones
Three Classes of Hormones
1. Polypeptides and protein hormones stored in secretory
vesicles until needed
+ Anterior & posterior pituitary hormones
+ Pancreas ( insulin & glucagon)
+ Parathyroid gland (parathyroid hormone)
2. Derivatives of the amino acid tyrosine
+ Thyroid ( thyroxine & triiodothyronine)
+ Adrenal medullae ( Epinephrine & norepinephrine)
3. Steroids synthesized from cholesterol and are not stored
+ Adrenal cortex (cortisol & aldosterone)
+ Ovaries ( estrogen & progesterone)
+ Testes ( testosterone)

Cell of origin Chemical messenger Site of release

Body function
„ Neurons Neurotransmitters Synaptic junctions

nerve cell function

„ Glands Endocrine hormones Blood

cell function in the body

„ Neurons Neuroendocrine hormones Blood

cell function in the body

„ Cells Paracrine hormones Extracellular fluid

function of different type

of cells
„ Cells Autocrine hormones Extracellular fluid

cell function of same cells

 Classes of Hormone receptors

Classes of Hormones
Type Example Synthetic path Mode of action
Peptide Insulin, Proteolytic Plasma
glucagon, processing of
pit, hypoth prohormone membrane

Catecholamine Epinephrine From tyrosine receptors;

2nd
messengers

Steroid Testosterone From cholesterol Nuclear

cortisol
Vitamin D 1,25-Dihydroxy- From cholesterol receptors;
cholecalciferol
Thyroid Tetraiiodothyronine From tyrosine transcriptional
(T4) in regulation
Triiodothyronine thyroglobulin
(T3)

Pit=pituitary; hypoth=hypothalamic
Neuroendocrine
system

Coordination of
Metabolism
 TSH FSH LH GH

Details of the hypothalamus-pituitary system. Signals from

connecting neurons stimulate the hypothalamus to secrete
releasing factors into a blood vessel that carries the
hormones directly to a capillary
network in the anterior pituitary.

Release of
hypothalamic
factors into
arterial blood Posterior pituitary hormones
are synthesized in neurons
arising in the hypothalamus,
transported
along axons to
nerve endings in
the posterior pituitary &
stored there until released
In response to each hypothalamic into the blood in response to
releasing factor, the anterior pituitary a neuronal signal.
releases the appropriate hormone
into the general circulation.

Release of posterior pituitary hormones

Release of ( vasopressin, oxytocin)
anterior pituitary
hormones
Veins carry hormones to systemic blood
( tropins)
Cascade of hormone release following central nervous system input to the hypothalamus

In each endocrine tissue along the

pathway, a stimulus from the level
above is received, amplified, &
transduced into the release of the next
hormone in the cascade. The cascade is
sensitive to regulation at several levels
through feedback inhibition by the
ultimate hormone. The product
therefore regulates its own production.

Synthesis & Secretion of peptide hormones

I. Gene Coding:
+ Proopiomelanocortin (POMC) is
precursor for eight hormones

+ Oxytocin & Vasopressin encoded on

separate genes together with their respective
neurophysin proteins
II. Synthesis on ribosomes in the form of
prohormones
III. Packaging into secretory vesicles with
proteolytic cleavage
Prepro-
hormone

Prohormone

Hormone

Products in the corticotrophs of the

anterior pituitary under control of CRH

Products
in the

hypothalamus
Amino Acid
-Derived
Hormones

Biosynthesis

Packaging

& Release of Epinephrine

in adrenal medulla chromaffin cell

 Neurosecretory granules contain epinephrine, dopamine β-
hydroxylase, ATP, Met or Leu-enkephalin, & larger enkephalin-
containing peptides or norepinephrine in place of epinephrine.
EP & NEP are often stored in different granules.

PNMT=phenylethanolamine methyl transferase; EP, epinephrine; NEP,

norepinephrine.Tyr,tyrosine.

peroxidase
Synthesis & Structure
of Thyroid
hormones

Thyroxine Triiodothyronine coupling

T4 T3
Hormone secretion after splitting of
the amines from thyroglobulin

Free hormone release into blood

Combination with plasma proteins

mostly with thyroxin-binding globulin
Cellular mechanisms for T4 & T3 release into the blood. Secretion requires endocytosis of thyroglobulin & subsequent
proteolysis. DIT & MIT are deiodinated & the released iodide ions are reutilized for hormone synthesis.
2.
Oxidation
of the
iodide ion

1. Iodide
pump( Iodide
3.
trapping)
Na+/I-
symporter 4.
(NIS)

5.
Release of T4 & T3
from the thyroid
gland
MIT= monoiodotyrosine; DIT= diiodotyrosine; TG = thyroglobulin which is a large glycoprotein stored in lumen of thyroid follicles

Cyclic hormonal Systems

„ Diurnal variation in the secretion of cortisol

from the adrenal cortex is regulated by the
sleep/wake cycle.
„ The female ovarian cycle operates on a
cyclic basis dictated by the central nervous
system
„ These are examples of chronotropic
control of hormone secretion
 Circadian Rhythm of ACTH & Cortisol Secretion

Time elapsed (hour)

Membrane Hormone Receptors

 Effect of TSH on Secretion of
Thyroid Hormone
TSH stimulates all steps in synthesis & secretion of T3 &
T4. These are mediated by its binding to TSH receptors
located on basal membrane of thyroid epithelial cells,
elevation of cAMP levels, & subsequent cascade of
phosphorylation reactions.

phosphodiesterase

5’AMP

TSH Model of TSH receptor

2. The specificity of receptor
recognition is imparted by the β-
subunit with a unique structure
for each hormone.

1. Thyrotropin (TSH), luteinizing

hormone (LH), & follicle-stimulating
hormone (FSH) each contain an α-&
a β-subunit. The α-subunits for all 3
hormones are identical.
3. The TSH-receptor complex
stimulates adenylate cyclase
& the phosphatidylinositol
pathway
 Internalization of Receptors
„ Many types of cell membrane hormone-
receptor complexes are internalized by
endocytosis
„ Endocytosis renders a cell less responsive to
hormone, since it reduces the number of cell
surface receptors receptor down-
regulation with a decrease in hormone sensitivity

For endocytosis to occur, the polypeptide receptor complex enters coated pits, which are
indentations of the plasma membrane into the cytosol. The coated pits pinch off from the
membrane to form coated vesicles, which shed their coats, fuse with each other, & form
vesicles called receptosomes. The receptors & ligands on the inside of
these receptosomes have different fates.

Clathrin directs internalization of

Hormone-Receptor Complexes from
plasma membrane

2. Coated vesicles fuse with lysosomes, which

contain proteolytic enzymes that degrade
both receptor & hormone.
3. The hormone
is only degraded
with the receptor
returning intact
to the membrane
1. Receptors may be recycled to cell surface
following fusion with the Golgi apparatus.

4. The receptor may

also be concentrated
in coated pits in the
absence of exogenous
ligand & cycle in &
out of the cell in a
constitutive, non-
ligand-dependent
manner.

A diagrammatic summary
of endocytosis in cells
 Intracellular Hormone Cascade:

Protein Kinases

Receptor: Transduction Through Tyrosine Kinase

The α-subunits of the insulin receptor are located extracellularly & are the insulin-binding sites. Ligand binding
induces autophosphorylation of tyrosine residues located in the cytoplasmic portions of β-subunits. This
autophosphorylation facilitates binding of cytosolic substrate proteins, such as insulin receptor substrate-1 (IRS-1).
Upon hormone binding, insulin receptor undergoes tyrosine autophosphorylation & kinase activation. It
phophorylates intracellular substrates IRS-1, Shc, & Cbl, which associate with SH2-containing proteins like p85 &
Grb2. Formation of the IRS-1/p85 complex activates PI(3-) kinase;
Insulin α-subunit & IRS-1/Grb2 complex activates MAP kinase.
4
UT
Insulin β-subunit GL
IRS-1
Receptor Tyrosine
Kinase p85 PI-3K
PO4
raf s Phosphorylated IRS-1
syp Grb2
ra OS acts as a docking protein
for proteins mediating glu
C S insulin action.

SH KK
Protein Lipid Glycogen
ME Synthesis Synthesis Synthesis
K
ME The net responses to insulin include short-term
metabolic effects, such as a rapid increase in the
PK uptake of glucose, & longer-term effects on cellular
MA differentiation & growth.

Transcription Factors

PI-3K, phosphatidylinositol-3-OH kinase; MAP kinase, mitogen-activated protein kinase; GLUT4, glucose transporter.

2
3

1
1
5
4

6 7

Regulation of Secretion of LH & FSH by protein kinase C

GnRH, gonadotropin-releasing hormone; FSH, follicle-stimulating hormone; LH, luteinizing hormone; DAG, diacylglycerol.
Examples of Hormones that operate Through the protein kinase A
Hormone Location of Action
CRH (corticotropin-releasing hormone) Corticotrope of anterior pituitary

TSH ( thyrotropin) Thyroid follicle

Leydig cell of testis
LH ( luteinizing hormone)
Mature follicle at ovulation & corpus luteum

FSH ( follicle-stimulating hormone) Sertoli cell of semineferous tubule &

ovarian follicle

ACTH ( adrenocorticotropic hormone)

Inner layers of cells of adrenal cortex

AVP ( arginine vasopressin)

Kidney distal tubular cell

Norepinephrine/epinephrine β-Receptor: expressed in various tissues &

cell types

Polypeptide Hormones that ↑ phosphatidylinositol pathway & ↑ Protein Kinase C

Hormone Location of Action
TRH (thyrotropin-releasing hormone) Thyrotrope of anterior pituitary releasing TSH

Gonadotrope of anterior pituitary releasing LH & FSH

GnRH ( gonadotropin-releasing hormone)

Corticotrope of anterior pituitary: assists CRH in

AVP ( arginine vasopressin)‡
releasing ACTH

TSH ( thyrotropin)§ Thyroid follicle: releasing thyroid hormones by↑ in

phosphatidylinositol as well as ↑ in protein kinase A

Angiotensin II/III Zona glomerulosa cell of adrenal cortex:

releases aldosterone

Epinephrine Smooth muscle cells that express α1-

receptors

§ TSH uses more than one intracellular second messenger system to transmit its signal
‡ Although AVP activates protein kinase A in renal cells, this same hormone activates protein
kinase C in other target cells.
 Model for signal transduction by ANF receptor
The receptor for ANF is a transmembrane protein whose
cytoplasmic C-terminal domain has guanylate cyclase
activity with extracellular N-terminal domain that binds
ANF.
The guanylate cyclase domain is in a
highly phosphorylated state under
normal conditions. Binding of ANF ↑
enzyme activity & dephosphorylation
of the guanylate cyclase domain.

The cGMP that is produced

activates protein kinase G,
which then phosphorylates
other cellular proteins
involved in this pathway.

ANF, atrial natriuretic factor Activity of ANF Protein Kinase G

Steroid Hormones
 Steroid Nucleus
Steroid hormones are divided into 2 classes: the
sex & progestational hormones & the
adrenocortical hormones. They are synthesized
in the gonads ( ovaries & testes ) & adrenal
cortex from cholesterol through pregnenolone
as intermediate.Their structure is based on the
cyclopentanoperhydrophenanthrene nucleus
with the numbering of this ring system & the
lettering presented here.

cortisol
Adrenal cortex aldosterone
androgens
estrogen
Gonads progesterone
testosterone

Glucocorticoids

Mineralocorticoids

C21-carbon compound C21-carbon compound

Sex Steroids

C19-carbon compound C18-carbon compound

The adrenocortical cell takes up the LDL particle via the LDL receptor & receptor-mediated endocytosis into clathrin-coated vesicles.
Lysosomal hydrolases act on the cholesteryl esters to release free cholesterol. The cholesterol nucleus, whether taken up or synthesized de
novo, subsequently undergoes a series of reactions that culminate in the formation of pregnenolone.
Unlike peptide & amine hormones, steroid hormones are not stored in secretory vesicles before their secretion.
Divided into 2 classes
Steroid-secreting cells are capable of ↑ secretion of steroids within several hours.

Steroid
Hormones

Steroid hormones mediate all their

actions on target tissues by regulating
gene transcription as a result, the
response of target tissues to steroids
occurs over hours to days.

Steroid hormones are poorly soluble in water. On their release into the
circulation, they associate with specific binding proteins & are transported to
target tissues through the circulatory system

Upon hormone binding to the receptor ( i.e ACTH for cortisol synthesis):
* ↑ adenylate cyclase via a stimulatory G protein & phosphatidylinositol cycle (PI)
* ↑ IP3 ↑ cytosolic Ca++ & cAMP ↑protein kinase A

↑Ca++ & protein phosphorylation overcome the rate-limiting steps in steroid synthesis:
1. Availability of free cholesterol from cholesteryl esters stored in vesicles
2. Transport of cholesterol to inner mitochondrial membrane by the StAR protein
3. Side-chain cleavage reaction

2 3 4 CYP11B2 5 Angiotensin II
Regulation

CYP11B1 ACTH
7 Regulation

StAR, steroidogenic acute regulatory protein

Biosynthetic pathways for adrenal steroid production

 Transport & Metabolism of Steroid Hormones

„ Cortisol bound to corticosteroid–binding globulin (CBG)

(80%) (Transcortin)↑ in pregnancy & estrogen intake ↑ total cortisol

~ 15% bound to albumin

~ 5-10% of cortisol is free
Slow metabolism with a ½ life of 60-70mns
„ Vs bound Aldosterone (50%)

Rapid metabolism with a ½ life of 20 mns

„ Liver principal site for steroid metabolism

Enzymatic reactions involved ↓ their biological activity &↑ their solubility in water with excretion in urine.
Conjugation also ↑ water solubility with glucuronides & sulfates most common conjugates.

■ Sex-hormone binding globulin binds testosterone & estradiol

Overview of Hormonal

Stimulation of

Steroid Hormone

Biosynthesis
 1. Nature of the hormone depends on the cell type & receptor
(ACTH for cortisol synthesis, FSH for estradiol synthesis, LH for
testosterone synthesis)

2. This activates adenylate cyclase via a

stimulatory G protein or may stimulate a Ca
channel directly or indirectly by activating
3. If PI is ↑, IP3 ↑ cytosol Ca++ levels
from ER store. the PI cycle.

4. cAMP ↑ protein kinase A, which via

phosphorylation, as well as induction
of the StAR protein, result in ↑ side-
chain cleavage & steroid
synthesis.These reactions overcome
the rate-limiting steps in steroid
biosynthesis, & more steroid is
synthesized & secreted.

Cellular Mechanism of Cortisol Action

Step8: alteration in the metabolic
activity of the target cell
Step1
Dissociation
Step 2

Step5:
Translocation Step7: synthesis
of new proteins

Step 3: binding

Step4: activation
of hormone-
receptor complex
to DNA-binding Step6: binding of activated
form hormone-receptor complex
to specific response
elements within DNA
3 Major Functional Domains of Steroid Receptor Proteins
Located at the C terminus, the steroid-binding domain has
30-60% homology with the ligand-binding domains of other
steroid receptors. This domain may also be involved in the
binding of a dimer of the 90-KDa heat shock protein which
has 2 functions: 1. It maintains ligand-binding domain in
optimal conformation for steroid binding. 2. It prevents
unliganded receptor from binding to DNA.

To the left of this domain is a region that modifies transcription & contains a
nuclear localization signal which provides recognition for the nuclear pore. In
the center is the DNA-binding domain which shows 60-90% homology among
steroid receptors. This domain contains 2 zinc fingers that recognize specific
HREs & stabilize binding to these DNA sequences.
The N-terminal domain is variable among the steroid
receptors & contains the principal antigenic region &
a region that modulates transcriptional activation. These features are common
to all members of the steroid receptor superfamily: T3 (triiodothyronine); RA
(retinoic acid); D3 (dihydroxyvitamin D3); E2( estradiol); Cortisol; androgen;
progesterone; aldosterone.

DBD, DNA-binding domain; LBD, ligand-binding domain; HRE, hormone-response-element