Mechanism of Hormone Action

Norman Arie Prayogo, S.Pi,M.Si

Two systems control all physiologic processes

The nervous system exerts point-to-point

control through nerves. Nervous control is
electrical in nature and fast.

The endocrine system broadcasts its

hormonal messages to essentially all cells by
secretion into blood and extracellular fluid. It
requires a receiver to get the message. In the
case of endocrine messages, cells must bear
a receptor for the hormone being broadcast in
order to respond.
 Hormon

Semua senyawa yang dapat mengubah atau

mempengaruhi fungsi tubuh
Dibagi menjadi 3 : endokrin, parakrin dan autokrin
Berupa peptida, steroid, eukosanoat, vitamin D dll
Endokrin akan dibahas lebih lanjut
Neurocrine signaling - hormone signal
originates in a neuron and transported down
the axon for delivery into the bloodstream
Sistem Endokrin
 Regulation of pituitary hormone secretion by the
hypothalamus
secretion of posterior pituitary hormones: a typical
neuroendocrine regulatory mechanism
secretion of anterior pituitary hormones: a typical
endocrine regulatory mechanism
important releasing/inhibiting hormones produced
in hypothalamus:
- Corticotropin-releasing hormone (CRH)
- Thyrotropin-releasing hormone (TRH)
- Gonadotropin-releasing hormone (GnRH)
- Growth-hormone-releasing hormone (GHRH)
- Prolactin-releasing hormone (PRH)
- Prolactin-inhibitory hormone (PIH)
 Posterior pituitary hormones
Antidiuretic hormone (ADH, vasppressin)
- chemistry: 9 amino acid peptide, produced
primarily by supraoptic nucleus and small
amount by paraventricular nucleus
- actions:
1) water retention by the kidney
urine volume and ECF
2) vasoconstriction (in large amounts)
- regulation/stimuli:
1) blood (or ECF) osmolality/osmoreceptors
2) blood volume
 3) others: alcohol, nicotine, barbiturates, etc.
- abnormality: diabetes insipidus
Oxytocin
- chemistry: 9 amino acid peptide, produced
primarily by paraventricular nucleus and small
amount by supraoptic nucleus
- actions: 1) regulating breast milk release
2) contraction of pregnant uterus
- regulation/stimuli:
suckling by a nursing infant
crying sounds from a baby
fear and stress inhibit release
 anterior pituitary hormones
Growth hormone
Prolactin
Thyroid-stimulating hormone
Adrenocorticotrophic hormone
Folicle-stimulating hormone
Luteinizing hormone
Growth hormone (GH)
- chemistry:191 peptide
- actions: see Fig.13-7
- regulation/stimuli:
Growth hormone releasing hormone (GHRH)
Feedback Negatif
Feedback Positif
B. The Thyroid Hormones
The main hormones secreted by the thyroid gland are
iodinated derivatives of thyroxine (T4) and
triiodothyronine (T3).
Synthesis and secretion of thyroid hormones
 Regulation of thyroid synthesis
and secretion - hypothalamic-
pituitary-thyroid axis
Hypothalamus TRH
anterior pituitary TSH
thyroid T3 & T4 negative
feedback on the hypothalamus
and the anterior pituitary
C. The Adrenal Glands
Adrenal medulla
Adrenal cortex
Three specific zones and each produces a specific
class of steroid hormone
Zona glomerulosa - mineralocorticoids
Zona fasciculata - glucocorticoids
Zona reticularis - androgens
 glucocorticoids (cortisol)
- regulation of glucocorticoid secretion
 Hormones, Receptors and Target Cells
Most hormones circulate in blood, coming into contact with
essentially all cells.
A given hormone usually affects only a limited number of
cells, which are called target cells.
A target cell responds to a hormone because it bears
receptors for the hormone.
Receptor`s Recognition and Coupling Functions

Recognition
Hormones present at very low concentration (10-15 to 10-9
mol/L) that that of other molecules (10-5 to 10-3 mol/L).
Receptors have to distinguish different hormones (specific)
present in very small amounts (sensitive).

Coupling (signal transduction)

Receptor-effector coupling provides amplification of
hormonal response.
Plasma carrier proteins only bind hormone but do not
generate a signal.
Signal amplification
of epinephrine
Nonsteroid hormones
bind exclusively to
plasma membrane
receptors, which mediate
the cellular responses to
the hormone. Steroid
hormones exert their
effects either by binding
to plasma membrane
receptors or by diffusing
to the nucleus, where
they modulate
transcriptional events.
 Hormones and Receptors

Principle Mechanism of
Location of Receptor Classes of Hormones
Action

Generation of second
messengers which alter
Proteins and peptides,
Cell surface receptors the activity of other
catecholamines and
(plasma membrane) molecules - usually
eicosanoids
enzymes - within the
cell

Intracellular receptors Alter transcriptional

Steroids and thyroid
(cytoplasm and/or activity of responsive
hormones
nucleus) genes
Structure of Intracellular Receptors
All of these receptors are composed of a single polypeptide
chain that has, in the simplist analysis, three distinct
domains:

The amino-terminus: In most cases, this region is

involved in activating or stimulating transcription by
interacting with other components of the transcriptional
machinery. The sequence is highly variable among different
receptors.

DNA binding domain: Amino acids in this region are

responsible for binding of the receptor to specific
sequences of DNA.

The carboxy-terminus or ligand-binding domain: This is

the region that binds hormone.
In the case of glucocorticoid receptors, there are 2
additional domains

Transport domain : responsible for translocation of the

receptor from cytoplasm to nucleus

Other domain that bind other protein (heat shock

protein) in the absence of ligand.
The primary structures of nuclear steroid
and thyroid receptor proteins
Structure of Cell Surface Receptors
Cell surface receptors are integral membrane proteins have regions
that contribute to three basic domains:

Extracellular domains: Some of the residues exposed to the outside of

the cell interact with and bind the hormone - another term for these
regions is the ligand-binding domain.

Transmembrane domains: Hydrophobic stretches of amino acids are

"comfortable" in the lipid bilayer and serve to anchor the receptor in the
membrane.

Cytoplasmic or intracellular domains: Tails or loops of the receptor

that are within the cytoplasm react to hormone binding by interacting in
some way with other molecules, leading to generation of second
messengers. Cytoplasmic residues of the receptor are thus the effector
region of the molecule.
All receptors that mediate transmembrane signaling processes
fall into three categories of receptor superfamilies:

1. The 7-transmembrane segment (7-TMS) receptors are integral

membrane proteins with seven transmembrane (helical) segments, an
extracellular recognition site for ligands, and an intracellular
recognition site for a GTP-binding protein (see following discussion).

2. The single-transmembrane segment (1-TMS) catalytic

receptors are proteins with only a single transmembrane segment
and substantial globular domains on both the extracellular and
intracellular faces of the membrane. The extracellular domain in the
ligand recognition site and the intracellular catalytic domain is either a
tyrosine kinase or a guanylyl cyclase.

3. Oligomeric ion channels consist of associations of protein

subunits, each of which contains several transmembrane segments.
These oligomeric structures are ligand-gated ion channels. Binding
of the specific ligand typically opens the ion channel. The ligands for
these ion channels are neurotransmitters.
Classification of hormones by mechanism of action

I. Hormones that bind to intracellular receptors

androgens, calcitriol, estrogen, gluco- and mineralocorticoid,
progestins, retinoic acids, thyroid hormones

II. Hormones that bind to extracellular receptors

A. The second messenger is cAMP
alpha2- and beta adrenergic catecholamine, ACTH,
ADH, angiotensin II, calcitonin, hCG, CRH, FSH,
glucagon, LH, MSH, PTH, TSH, somatostatin

B. The second messenger is cGMP

ANF, NO
C. The second messenger is calcium or
phosphatidylinositol or both
acetylcholine, alpha1-adrenergic catecholamine,
angiotensin II, ADH, cholecystokinin, gastrin, GnRH,
oxytocin, PDGF, substance P, TRH

D. The second messenger is a kinase or phosphatase

cascade
Chorionic somatomammotropin, EGF, erythropoietin,
FGF, GH, insulin, IGF-I, IGF-II, NGF, PDGF, prolactin
 General features of hormone classes

Group I Group II
Types Steroid, calcitriols, Polypeptides, proteins,
iodothyronins, retinoid glycoproteins,
catecholamines
Solubility Lipophilic Hydrophilis

Transport proteins Yes No

Plasma half-life Long (hours to days) Short (minutes)

Receptor Intracellular Plasma membrane

Mediator Receptor-hormone cAMP, cGMP, calcium,

complex phosphoinositols, kinase
cascade
GROUP I HORMONES
The hormones are transported to the cell target by its
transporter. It dissociate from the transporter and diffuses
through the plasma membrane and bind to the receptor.
The hormone-receptor complex binds to a specific region
of DNA called the hormone response elements (HRE).
This binding affect gene trancription and the production of
mRNAs, the amount of specific protein are chenged and
metabolic processes are influenced.
Another effect is DNA replication which will induce cell
proliferation.
HRE located in the 5 region of promoter element.
A model for steroid hormone action in target cells. The hormone
(for example, estrogen) dissociates from plasma proteins, diffuses
into the cell, and binds to receptor proteins. The active hormone-
receptor complex migrates to the nucleus, where it interacts with
DNA or transcription factors or both.
GROUP II HORMONES
The hormones are water soluble, no transport protein, have
membrane receptor and use intracellular messenger.

II A. cAMP
3,5-adenylic acid, derived from ATP through the action of
enzyme adenylyl cyclase
The intacellular level of cAMP increase or decrease by
various hormone.
 Subclassification of Group II A Hormones

Stimulate (Hs) Inhibit (Hi)

ACTH, ADH, Beta-adrenergic Acetylcholin, alpha2
Calcitonin, CRH, FSH, adrenergic, angiotensin II,
glucagon, hCG, LH, MSH, somatostatin
PTH, TSH
Cyclic AMP is synthesized by membrane-bound adenylyl
cyclase and degraded by soluble phosphodiesterase.
The mediator of cAMP formation (7-TMS) is GTP-binding
protein (G-protein)
Activation of Protein Kinase A (cAMP-
dependent Protein Kinase) by cAMP
 Enzymes Regulated by Protein Kinase A

Enzyme Pathway
Glycogen synthase Glycogen synthesis
Phosphorylase b kinase Glycogen breakdown
Hormone-sensitive lipase Lipid metabolism
Pyruvate dehydrogenase Pyruvate to acetyl-CoA
Tyrosine hydroxylase Dopamine synthesis
G Proteins
G proteins are heterotrimers consisting of - (45 to 47 kD), - (35 kD),
and - (7 to 9 kD) subunits. The -subunit binds GDP or GTP and has
an intrinsic, slow GTPase activity.

The G complex in the unactivated state has GDP at the nucleotide

site. Binding of hormone to receptor stimulates a rapid exchange of
GTP for GDP on G . The binding of GTP causes G to dissociate
from G and to associate with an effector protein such as adenylyl
cyclase. Binding of G (GTP) activates adenylyl cyclase. The adenylyl
cyclase actively synthesizes cAMP as long as G (GTP) remains bound
to it.

However, the intrinsic GTPase activity of G eventually hydrolyzes

GTP to GDP, leading to dissociation of G (GDP) from adenylyl cyclase
and reassociation with the G dimer, regenerating the inactive
heterotrimeric G complex.
Adenylyl cyclase activity is modulated by the interplay of
stimulatory (Gs) and inhibitory (Gi) G proteins.
 Cholera toxin

Vibrio cholerae, the Gram-negative bacterium that causes cholera,

induces severe diarrhea in its victims, leading to death if the fluids are
not replenished.
Cholera toxin is an 87-kD protein consisting of an A subunit and five B
subunits. The B subunits act in host-cell recognition. The A subunit
consists of A1 and A2 peptides linked by a disulfide bond. The 22-kD
A1 peptide catalyzes the ADP-ribosylation of Arg201 in the of Gs
using NAD+ as a substrate. ADP-ribosylation strongly inhibits the
GTPase activity of Gs, effectively trapping Gs in the activated state
and causing prolonged activation of adenylyl cyclase.
Elevated levels of cyclic AMP in turn cause intestinal epithelial cells to
secrete high volumes of fluid. Both the bacteria and the cholera toxin
remain localized in the intestines through the course of the disease, but
if fluids are actively replaced, the bodys immune system eventually
gains control and destroys the bacteria.
Pertussis toxin Causes ADP-Ribosylation of Gi

ADP-ribosylation of a Cys residue on Gi is catalyzed

by pertussis toxin, a product of Bordetella pertussis,
the bacterium that causes whooping cough.
Pertussis toxin is a 110-kD hexameric protein (a 28-kD
A subunit and five B-like subunits.) In this case, the
ADP-ribosylation inhibits exchange of GDP for GTP,
thereby preventing Gi from inhibiting adenylyl
cyclase. Pertussis, in contrast to cholera, is a systemic
infection, and the breakdown of adenylyl cyclase
regulation is felt by tissues throughout the body.
 II B. cGMP

Another cellular second messenger,

guanosine 3',5'-cyclic
monophosphate (cGMP), is
formed from GTP by guanylyl
cyclase. Membrane-bound
guanylyl cyclases constitute a
second class of single-TMS
receptors with an extracellular
hormone-binding domain; a single,
a-helical transmembrane segment;
and an intracellular catalytic
domain. cGMP is hydrolyzed by
cGMP phosphodiesterase
A variety of peptides act to stimulate the membrane-
bound guanylyl cyclases, including atrial natriuretic
peptide (ANP), which regulates body fluid
homeostasis and cardiovascular function; and nitric
oxide (NO) which links cGMP to vasodilatation.

cGMP increase by activating guanylyl cyclase

(nitrprusside, nitrogliserin, sodium nitrite) or inhibit
cGMP phophodiesterase (sildenafil/Viagra)

Protein kinase G (cGMP-dependent protein kinase)

phosphorylates ser and thr residues in target
proteins. Activation of protein kinase G is similar
with that of protein kinase A.
 II C. Calcium and PI

Calcium ion is an important intracellular signal. Binding of

certain hormones and signal molecules to plasma
membrane receptors can cause transient increases in
cytoplasmic Ca2+ levels, which in turn can activate a
wide variety of enzymatic processes, including smooth
muscle contraction, exocytosis, and glycogen
metabolism.
Calmodulin is calcium dependent regulatory protein.
Calmodulin has four binding sites for calcium. Interaction
of calcium and calmodulin activates Ca/calmodulin-
dependent protein kinase (CaM kinase), then activates
other molecules (ion transporters, enzymes , e.g. :
adenylyl cyclase, glycogen synthase, pyruvate kinase).
Cytoplasmic [Ca2+]
increases occur via the
opening of Ca2+ channels
in the membranes of
calciosomes, the
endoplasmic reticulum,
and the plasma
membrane.
IP3 (inositol 1,4,5-P3) mediates Ca2+-induced Ca2+
release. Binding of IP3 to the ER IP3 receptor opens
ER Ca2+ channels. Flow of Ca2+ through these
channels induces a conformational change that opens
plasma membrane Ca2+ channels.
 The pathways of phosphoinositide
biosynthesis and metabolism

Release Calcium from ER

IP3 has a cellular half-life of only a few seconds. It is
rapidly processed along two principal paths: (a) it can
be catabolized by a series of phosphatases to yield
inositol-1,4-bisphosphate, inositol-4-phosphate, and
free myo-inositol, which can be subsequently
reincorporated into new inositol phospholipids; or (b) it
can be phosphorylated to yield inositol-1,3,4,5-
tetraphosphate, which then undergoes a complex
series of phosphorylations and dephosphorylations to
at least six other inositol phosphate compounds.
Calcium-dependent protein kinase (Protein kinase C)

PKC is specifically activated by two intracellular second

messengers: diacylglycerol and Ca2+ (the C in PKC stands for
Ca2+). Because Ca2+ levels increase in the cell in response to
IP3, the activation of PKC depends upon both of the second
messengers released by hydrolysis of PIP2.
PKC is a cellular transducer, translating the hormonal message
and the signals of second messengers into the protein
phosphorylation events that control growth and development.
At low levels of Ca2+ and in the absence of DAG, protein kinase C
is inactive and is a soluble protein in the cytoplasm. In this state, the
pseudosubstrate domain occupies the substrate-binding site,
keeping the enzyme inactive.
DAG binding causes conformation changes that dissociate the
pseudosubstrate domain from the substrate-binding site and
increase the affinity of the enzyme for Ca2+ and lipid, causing
protein kinase C domain C1 to bind to the cytoplasmic surface of
the plasma membrane, whereupon the kinase becomes active.
Control of enzymatic activity by the insertion of a pseudosubstrate
domain into the active site has been referred to as intrasteric
control, in contrast to allosteric control, in which an enzyme
regulator with a structure unrelated to the substrate binds at a site
separate from the active site.
Activation of protein kinase C
II C. Protein kinase cascade

EGF receptor contain

intrinsic tyrosine kinase
activity. Bind EGF to
receptor activates
tyrosine kinase and
subsequently
phosphorilates other
molecules (enzymes)
Insulin transmit signals by several kinase cascades
 Hormone Actions

Activation of enzymes and other dynamic molecules: Most

enzymes shuttle between conformational states that are catalytically
active versus inactive, on versus off. Many hormones affect their target
cells by inducing such transitions, usually causing an activation of one
of more enzymes. Because enzymes are catalytic and often serve to
activate additional enzymes, a seemingly small change induced by
hormone-receptor binding can lead to widespread consequences
within the cell.

Modulation of gene expression: Stimulating transcription of a group

of genes clearly can alter a cell's phenotype by leading to a burst of
synthesis of new proteins. Similarly, if transcription of a group of
previously active genes is shut off, the corresponding proteins will
soon disappear from the cell.