HORMONES

(mainly Chapter 23 Lehninger)

 HORMONES AND HORMONAL REGULATION

• Hormonal regulation
involves a hierarchy of
cell types acting on each
other either to stimulate
or to modulate the
release and action of a
hormone.

• Secretion of hormones
from endocrine cells is
stimulated by chemical
signals from regulatory
cells that occupy a higher
position in this hierarchy.
 ENDOCRINE AND NEUROENDOCRINE SYSTEMS

• Nerve (electric) pulse originates in cell

bodies and they are rapidly sent, also
for long distance, to axon terminals,
where a neurotransmitter is released
to the target cell that is within a
distance of few µm.

• In endocrine system, hormones are

secreted to blood flux up to the target
tissue that can be more than 1 m
distant from the origin point
THE HIERARCHICAL NATURE OF HORMONE
ACTION
 THE HIERARCHICAL NATURE OF HORMONE
ACTION
• The hierarchical nature of hormone action can be summarized as follows:

1. Hormonal action is controlled ultimately by the central nervous system, which transmits signals to the
hypothalamus. It responds by producing factors that either stimulate (called releasing factors) or inhibit
the release of hormones from the pituitary.

2. Pituitary hormones do one of the following:

a. They stimulate other endocrine glands, each of which releases a hormone that acts on a target
tissue and elicits a specific metabolic response.

b. Alternatively, they act directly on a target tissue. The action of a hormone sets in motion events
that ultimately limit that action.

• Some pituitary hormones stimulate target tissue directly. For example, prolactin stimulates mammary
glands to produce milk.

• Most pituitary hormones act on endocrine glands that occupy an intermediate, or secondary, position
in the hierarchy, stimulating them to produce hormones that exert the ultimate actions on target
tissues. Pituitary hormones that act on other endocrine glands are called tropic hormones or
tropins.
 GENERAL MECHANISMS OF HORMONES ACTION
• Peptide and ammine hormones action is more rapid than the one generated by steroid and thyroid hormones.

Intracellular effects of hormones action are:

1. A second messenger (cAMP, cGMP for

example) is generated and allosterically
regulates one or more enzymes.

2. A tyrosine-kinase receptor is activated by the

extracellular hormone.

3. A variation in the membrane potential causes

the opening/closing of a ion channel controlled
by the hormone.

4. An adhesion receptor on cell surface transfer

the information from extracellular matrix to the
cytoskeleton

5. A steroid or a steroid-similar molecule causes a

variation at trascriptional level of one or more
genes through hormonal nuclear receptors.
GENERAL MECHANISMS OF HORMONES ACTION
 GENERAL MECHANISMS OF HORMONES ACTION

• The action of a hormone is self-limiting because of

the existence of feedback loops, in which secretion
of a hormone sets in motion a series of events that
leads to inhibition of that secretion.
GENERAL MECHANISMS OF HORMONES ACTION
 GENERAL MECHANISMS OF HORMONES ACTION

Scatchard Analysis

• The receptor-ligand binding, like the enzyme-

substrate binding, depends on the concentration of
the interacting components

• The resulting equilibrium can be described in term

of Ka (the association constant) or Kd (the
dissociation constant)
HORMONES CLASSIFICATION
 PEPTIDES
• Nearly all peptide hormones are synthesized as inactive
precursors and then converted to active hormones by
proteolytic processing.

• Insulin contains two polypeptide chains, of 21 and 30

residues, with two inter-chain disulfide bridges and one
intra-chain bridge.

• The first product of translation of the insulin gene is a

105-residue polypeptide called pre-proinsulin.

• Cleavage from pre-proinsulin of a 24-residue N-terminal

"signal sequence" yields proinsulin, an 81-residue
polypeptide.

• Proinsulin then undergoes folding, disulfide bond

formation, and cleavage to give the two polypeptide
chains of the active hormone, insulin.

• The signal sequence that is eventually cleaved from

preproinsulin to form proinsulin is needed to transport the
protein through membranes.

• All known polypeptide hormones are synthesized in "pre-

pro" form, with a signal sequence and additional
sequence(s) that are cleaved out during maturation of the
hormone.
 PEPTIDES
• A particularly interesting case is that seen
when a single polypeptide sequence
contains two or more distinct hormones.

• The most complex example is a pituitary

multihormone precursor that contains
sequences for β- and γ-lipotropin, α-,β -,
and γ-melanocyte-stimulating hormone
(MSH), endorphin, enkephalin, and ACTH.

• This precursor, called pro-opiomelanocortin,

derives its name from its role as precursor
to endogenous opiates, melanocyte-
stimulating hormone, and corticotropin.

• A remarkable fact about pro-

opiomelanocortin is that it is cleaved at
different sites in different cells, so that
different cell types produce different
ensembles of hormones derived from this
one precursor.

• In the anterior pituitary, cleavage generates

ACTH and β-lipotropin, and further
processing in the central nervous system
yields endorphin and enkephalin, among
other products.
PEPTIDES

Endogenous opioid peptides

Met-enkephalin is Tyr-Gly-Gly-Phe-Met.

Leu-enkephalin has Tyr-Gly-Gly-Phe-Leu.

 PEPTIDES

• The C-term residue of these two hormones

is glycinammide.

• The amidation of small peptide hormones is

common

• These two hormones differ in two residues

(pink box) but they have completely
different effects

• Oxytocin act on smooth muscle of the

uterus and mammary gland

• Vasopressin increases water adsorption in

kidney and induces blood vessels
constriction
CATECHOLAMINES

Catechol
 Catecholamines catabolism
• Catabolic process - in the liver
• Catechol-O-methyltransferase
• Use of the new S-adenosylmethionine (SAM) which is converted into S-adenosylhomocysteine
(SAO) with methylation of the OH in the aromatic ring
• met-adrenaline is formed (epinephrine O-methylated) that
• or is attacked by the MAO = monoamine oxidase, giving methyl-amine and aldehyde 3-methoxy-4-
hydroxymandelic which is dehydrogenated to acid 3-methoxy-4-idrossimandelic
• or it is combined with ac. glucuronic or ac. sulfuric
• all these forms are excreted in the urine

Catechol-O-
methyltransferase

Conjugation
MAO
through transferase

dehydrogenase
 EICOSANOIDS
• Eicosanoids are a class of lipids that
include the prostaglandins, thromboxanes,
and leukotrienes.

• Eicosanoids derive their name from their

common origin, that is, from C20
polyunsaturated fatty acids, the
eicosaenoic acids, particularly arachidonic
acid (all-cis-5,8,11,14-eicosatetraenoic
acid).
 EICOSANOIDS
• Eicosanoids exert short range effects in neighboring cells and tissues

• Eicosanoids (like prostaglandin) exert specific physiological effects on target cells, like hormones

• However, eicosanoids are distinct from most hormones in that they act locally, near their sites of
synthesis, and they are catabolized extremely rapidly. Thus, eicosanoids are considered to be locally
acting hormones.

Summary of biosynthetic routes to the major Structures of the major prostaglandins and
prostaglandins and thromboxane A2. thromboxane
 PROSTAGLANDINES AND THROMBOXANS
• Every prostaglandin contains 20 carbon atoms, including a 5-carbon ring.
• Prostaglandins are powerful locally acting vasodilators and inhibit the aggregation of blood platelets.
• Through their role in vasodilation, prostaglandins are also involved in inflammation.
• They include prostacyclins
• Specific prostaglandins are named with a letter (which indicates the type of ring structure) followed by
a number (which indicates the number of double bonds in the hydrocarbon structure).

PGE1 PGI2
(prostacyclin)

• Thromboxane is a member of the family of lipids known as eicosanoids. The two major thromboxanes
are thromboxane A2 and thromboxane B2.
• The distinguishing feature of thromboxanes is a 6-membered ether-containing ring.
• Thromboxane is named for its role in clot formation (thrombosis).
• Thromboxane is a vasoconstrictor and a potent hypertensive agent, and it facilitates platelet aggregation.

A2 B2
 Cycloxygenase (COX)
• In mammals, two isoforms of COX exist (COX1 and
COX2).

• They have similar sequences (60-65% identity), similar

reaction mechanisms but different roles

• COX1 is responsible for the synthesis of

prostaglandins that regulates secretion of the gastric
mucosa

• COX2 is responsible for the biosynthesis of

prostaglandins that mediate inflammation, pain and
fever

• Aspirin, an inhibitor of both COX1 and COX2, was

introduced into the market in 1899

• The drug was able to acetylate a serine residue

blocking the active site

• Ibuprofen (a FANS) inhibits both COX1 and COX2

 Cycloxygenase (COX)
• COX1 inhibition has major side effects such as stomach irritation

• In 90s, the crystal structure resolution of COX1 gave the opportunity to design inhibitors more specific for
COX2 for pain therapy

• Three drugs were designed and introduced in the market (rofecobix, valdecobix and colecobix)

• Different studies showed a correlation between the use of these drugs and heart attack and stroke so that
these drugs were withdrawn from the market

• Most probably, they alter the balance between prostacyclin and thromoboxanes, that control blood
coagulation
 Cycloxygenase (COX)
• COX is a heme-containing bifunctional protein that sequentially catalyzes two reactions.
• The first reaction involves cyclooxygenation of the endogenous substrate arachidonic acid to yield the
hydroperoxy endoperoxide PGG2.
• Subsequent reduction of the hydroperoxyl moiety of PGG2 results in formation of PGH2.
• The latter peroxidase reaction occurs in an adjacent, but spatially distinct, site within the COX catalytic
domain. Contains two separate active sites for prostaglandin synthase
• One side contains the cyclooxygenase active site
• The opposite side contains the peroxidase active site which is involved in activating the heme group
necessary for cyclooxygenase reaction
• Complex composed of identical dimers (2 cyclooxygenase sites and 2 peroxidase active sites)
• Each subunit has a carbon rich knob involved in anchoring the complex to the ER
• Knobs contain funnels to active sites responsible for guiding arachidonic acid from the ER to the enzyme

Crystallographic structures of ovine COX-1 (left) and murine COX-2 (right) homodimers. Functional domains: 1) membrane
binding domain (yellow); 2) dimerization domain (light green); catalytic domain (green) heme (red). The open cleft of the
peroxidase active site is observable at the top of each monomer. Glycosyl residues are not shown.
 EICOSANOIDS: LEUKOTRIENS
• Leukotrienes are eicosanoids that were originally isolated from leukocytes and contain three double
bonds, which explains how they were named.

• Leukotrienes are formed by a pathway independent of that of forming the prostaglandins and
thromboxanes.

• The pathway to leukotrienes starts by attack on arachidonate of a lipoxygenase, which adds O2 to C-5,
giving 5-hydroperoxyeicosatetraenoic acid (5-HPETE).
 VITAMIN D DERIVATIVE: CALCITRIOL
• The most abundant form of vitamin D is D3, called
cholcalciferol.

• Vitamin D is not technically a vitamin, because it is

not required in the diet.

• I t a r i s e s f r o m U V- p h o t o l y s i s o f 7 -
dehydrocholesterol, an intermediate in cholesterol
biosynthesis

• D3 undergoes two successive hydroxylations

catalyzed by mixed-function oxidases. The first
occurs at carbon 25 in liver.

• When calcium levels are low, hydroxylation occurs

at carbon 1, yielding the active form, 1,25(OH)D3,
which stimulates osteoblasts to take up calcium.

• In the intestine, 1,25(OH)D3 stimulates transcription

of a protein that stimulates calcium absorption into
the bloodstream.

• When calcium levels are adequate, hydroxylation

occurs instead at carbon 24, yielding the inactive
24,25(OH)D3 form.
VITAMIN D DERIVATIVE: CALCITRIOL
 RETINOID HORMONES
• Retinoids are potent hormones that regulate cell growth, survival and differentiation through nuclear
receptors

• Retinol is the pro-hormone and it is synthetised mainly in the liver from beta-carotene, whereas other
tissues convert retinol into retinoic acid
 THYROID HORMONES
• Thyroid hormones triiodothyronine (T3) and thyroxine (T4) are synthetised in thyroid gland from the protein precursor
tyroglobulin (Mr 660,000).

• Up tp 20 Tyr residues of the protein are enzymatically iodinated in thyroid gland

• Then, 2 residues of di-iodo-iodotyrosine condensate to form the precursor of thyroxine

• A proteolytic cleavage makes thyroxine available when required

• The condensation of a mono-iodo-tyrosine residue with a di-iodo-tyrosine give rise to triiodothyronine (T3)

• The condensation of two di-iodo-tyrosines give rise to thyroxine (T4)

 THYROID HORMONES

• Thyroid hormones triiodothyronine (T3)

and thyroxine (T4) stimulates energy
producing metabolism in liver and muscle

• They act through nuclear receptors

activating genes coding for catabolic key
enzymes
 IODINE METABOLISM
• Thyroid hormones are unique biological molecules in that they incorporate iodine in their
structure.

• Thus, adequate iodine intake (diet, water) is required for normal thyroid hormone
production.

• Dietary iodine is absorbed in the gastrointestinal tract, then taken up by the thyroid gland
(or removed from the body by the kidneys).

• The transport of iodide into follicular cells is dependent upon a Na+/I- cotransport system.

• Iodide taken up by the thyroid gland is oxidized by peroxidase in the lumen of the follicle:
I- I+
peroxidase

• Oxidized iodine can then be used in production of thyroid hormones.

 PRODUCTION OF THYROGLOBULIN
• Pituitary produces TSH, which binds to follicle cell receptors.
• The follicle cells of the thyroid produce thyroglobulin.
• Thyroglobulin is a very large glycoprotein.
• Thyroglobulin is released into the colloid space, where it’s tyrosine residues are iodinated
by I+.
• This results in tyrosine residues which have one or two iodines attached (monoiodotyrosine
or diiodotyrosine).

Figure 1: A) Biosynthesis of L-thyroxine

(T4) from thyroglobulin by thyroid
peroxidase (TPO) in the presence of H2O2
and iodide. The space-filling model
indicates the relative orientation of the
two iodinated phenyl rings in T4.

Angewandte Chemie Interna0onal Edi0on

Volume 54, Issue 37, pages 10833-10837, 24 JUL 2015 DOI:
10.1002/anie.201505281
THYROID HORMONES: SUMMARY

Häggström, Mikael. "Medical gallery of Mikael Häggström 2014". Wikiversity Journal of Medicine
1 (2). DOI:10.15347/wjm/2014.008. ISSN 20018762.
 TRANSPORT OF THYROID HORMONES

• Thyroid hormones are not very soluble in water (but are lipid-soluble).
• They leave the follicol cells through a monocarboxylate transporter (MCT)
• Thus, they are found in the circulation associated with binding proteins:
- Thyroid Hormone-Binding Globulin (TBG, ~70% of hormone)
- Pre-albumin (transthyretin), (~15%)
- Albumin (~15%)
• Less than 1% of thyroid hormone is found free in the circulation.
• Only free and albumin-bound thyroid hormone is biologically available to tissues.
• TBG-bound thyroid hormone does act as a reservoir of the hormone in the body
 ARGININE IS THE PRECURSOR OF THE SECOND
MESSENGER NO

GABA

NO

Hydroxyproline Creatine
phosphate

Collagen

• Nitric oxide is produced from arginine in an unusual five-electron oxidation that also yields citrulline
• The enzyme catalyzing the reaction, nitric oxide synthase, contains bound FMN, FAD, non-heme iron, and
tetrahydrobiopterin.
• Nitric oxide, is is a signal-transducing agent in the vasodilation of endothelial vascular cells and
underlying smooth muscle.
• It is also involved in signaling decreases in blood pressure, and inhibiting platelet aggregation.
• In the inflammatory and immune responses, an inducible form of nitric oxide synthase produces nitric oxide
at levels sufficient to be toxic to pathogenic organisms.
• It can act in neurotransmission in the central nervous system and stimulate erection of the penis.
• Nitric oxide is a gas so it can diffuse rapidly into neighboring cells and control their metabolism.
• It is also unstable, with a half-life of 1 to 5 seconds, so its effects are short-lived. In the cell, nitric oxide acts
primarily by stimulating cyclic GMP synthesis.
• The drug, Viagra, acts by inhibiting cyclic GMP breakdown, thereby prolonging the effect of nitric oxide.
 STEROIDS

SEE LECTURE 9