Mechanism of hormone action

Monday, 16 August 2021 7:35 AM

- In classic endocrine signalling, a hormone

secreted by an endocrine gland enters
systemic circulation and carries the signal
to a distant target tissue.
- In paracrine signalling, a hormone acts on
nearby cells without ever passing through
the systemic circulation.
- Finally, there is also autocrine regulation,
a hormone can act on the cell that itself
secretes the hormone.
(Some paracrine factors - interleukins,
PDGF; histamine secreted by ECL cells and
acting on the parietal cells of stomach,
somatostatin secreted by delta cells of
pancreas in regulation of insulin &
glucagon secretion.)
Paraneoplastic syndromes: Hormones are
produced by nonendocrine neoplastic
cells; the clinical syndromes that result
from secretion of these hormones are
called paraneoplastic syndromes. E.g.
Squamous cell carcinoma of lung is
associated with hypercalcemia, which
results from the secretion of a protein,
called PTH-related peptide, that mimics
the actions of PTH.
3 divisions or arms of the endocrine
system:
(I) Dedicated endocrine glands: They
synthesize and secrete the bioactive
hormones
(II) Non-endocrine organs (their primary
function is not endocrine): E.g., Kidney
synthesizes erythropoietin, GIT produces
GI hormones, heart synthesizes atrial
natriuretic peptide (ANP), etc.
(III) Cells in various organs that modify
inactive precursors or convert less active
hormones into more active hormones:
E.g., Formation of angiotensin II by the
action of ACE in the lung, formation of
vitamin D3 by hydroxylation reactions
successively in the liver and kidney.
General concepts of endocrine control:
Functions of hormones:
Most hormones have several different
target tissues. Many hormones (such as
thyroid, cortisol, etc.) have pleiotropic
actions (e.g., multiple phenotypic effects)
on numerous cell types.
Chemical nature/structure of the
hormones:
• Hormones may be peptides, metabolites
of single amino acids, or metabolites of
cholesterol-
a. Peptide hormones, proteins,
proteoglycans: The peptide hormone
families – (i) Insulin family: insulin, IGF-I &
IGF-II, relaxin. (ii) Glycoprotein family:
FSH, LH, TSH, hCG. (iii) Growth hormone
family: GH, prolactin, hPL. (iv) secretin
family: secretin, glucagon, VIP, GIP.
b. Derivatives of a single amino acid:
catecholamines and thyroid hormone are
derived from the amino acid tyrosine.
c. Steroids or metabolites of cholesterol:
synthesis of steroid hormones (from
cholesterol) necessitates a number of
enzymatic steps. Only very specialized
tissues are capable of the series of
enzymatic conversions that are necessary
to make active hormone from the starting
materials; e.g. adrenal cortex gland
synthesizes steroids – glucocorticoids and
mineralocorticoids
{* Peptide and protein hormones vary
greatly in size. E.g. Thyrotropin releasing
hormone (TRH) is a tripeptide; human
chorionic gonadotropin (hCG) has 243
amino acid residues.}
Hormone synthesis: general principles
a. Peptide hormones- Within the cell of the
endocrine gland the peptide molecule
prepared first is usually a larger precursor
called “preprohormone”; it is cleaved to
form “prohormone”; the further cleavage
of this peptide forms the final mature
hormone. (e.g. preproinsulin – to –
proinsulin – to – insulin). The hormone
may not be released immediately into the
circulation; it is stored in membrane-
bound secretory vesicles of endocrine
cells and released on demand. The
hormone is released by exocytosis
through the regulated secretory pathway.
That is, the peptide/protein hormones are
not secreted continuously; they are
secreted in response to a stimulus –
“stimulus-secretion coupling”.
b. Steroid hormones- are synthesized form
cholesterol; once formed they are
immediately released into circulation (not
stored)

c.
• Latent period for hormone action – It is
the time interval between the application
of stimulus and onset of a response. For
instance, oxytocin has among the shortest
latent periods; milk ejection occurs in a
few seconds. Thyroxine has one of the
longest latent periods; its metabolic
effects may take as long as 3 days to
begin.
Hormones can circulate either free or
bound to carrier proteins-
Most peptide hormones exist free in the
circulation. Steroid and thyroid hormones
circulate bound to plasma proteins. {IGF-I
and IGF-II are an exception to this rule: at
least six plasma proteins bind these
peptide growth factors.}
Forming a complex with a circulating
binding protein serves several functions:
(i) It provides the blood with a reservoir or
pool of the hormone, thus minimizing
minute-to-minute fluctuations in
hormone concentration. (ii) It extends the
half-life of the hormone in the circulation
(as only the free hormone is metabolized
by the liver or excreted by the kidney).
• Half-life in circulation: In general, peptide
hormones have shorter half-life in
circulation; steroids have longer half-life.
Epinephrine & norepinephrine have
among the shortest half-life (about 10-15
seconds). Thyroxine has a half-life of 7-8
days; 25(OH) vitamin D has a half-life of
15 days.
(The hormones bound to binding proteins
in plasma appear to be those whose
actions are chronic – in particular, those
involving induction of the synthesis of
new protein in target tissues. Hormones
that play a major acute role in the
regulation of body metabolism circulate
freely without associated binding
proteins.)
Ø Hormonal rhythms:
• Circadian (diurnal) rhythm: (24-hour
cycle) - cortisol, GH, prolactin
• Ultradian rhythms: (rhythms with a
periodicity of less than 24 hours) - LH,
FSH, testosterone
Mechanisms of hormone action
All the hormones exert their specific
actions on their target cells by binding to
specific receptors. The receptor proteins
can be divided into two broad categories:
(A) Cell membrane receptors – present on
the cell membrane, and (B) intracellular
receptors – present in cytoplasm or
nucleus of the target cell.
The protein/peptide hormones are not
lipid-soluble and cannot enter target cells
easily. Hence, receptors for these
hormones are situated on the cell
membranes. Steroid hormones are lipid-
soluble; they can cross the cell
membranes easily. Their receptors are
located inside the cells.
The protein/peptide hormones (the “first
messengers”) exert their effects without
entering the target cell. As they combine
with the membrane receptors, they
activate the so-called “second
messengers” within the cells. Second
messengers then activate the specific
enzymatic machinery within the cell to
produce the effect of the hormone.
A. THE CELL MEMBRANE RECEPTORS -
There are 2 major types of cell
membrane receptors for the
peptide/protein hormones ~
1. G protein-coupled receptors (GPCRs) –
These integral membrane receptor
proteins work through an intermediary;
the intermediary is a heterotrimeric GTP-
binding complex called G-protein. {GTP =
guanosine triphosphate). By acting
through this G- protein, they activate or
inactivate a separate membrane-
associated enzyme or channel.
2. Catalytic receptors - When activated by a
ligand/hormone, these integral
membrane receptor proteins are either
enzymes themselves or part of an
enzymatic complex.
{A category of ion-channel linked
receptors may also be included in the
‘membrane receptor’ family. It is
mostly utilized by neurotransmitters
such as ACh. These receptors help
regulate the intracellular concentration
of specific ions.}

1. G PROTEIN-COUPLED RECEPTORS: (GPCRs)

[Figure: A diagrammatic presentation of

the GPCR. When a hormone binds with
the GPCR, the a-subunit of the G-protein
dissociates from the bg subunits. It
activates an effector protein inside the
cell. For instance, activation of adenylyl
cyclase enzyme à converts cellular ATP
into cAMP which then acts as a second
messenger.]
- Receptors using G proteins comprise the
largest family (more than 1000 members)
of the hormone receptors. {G protein =
GTP-binding protein} The G protein-
coupled receptor (GPCR) family consists of
receptors that span the cell membrane 7
times, and are referred to as 7-helix
transmembrane receptors.
- These receptors interact with the
protein – the heterotrimeric G protein.
This G protein has 3 subunits: α, β, and γ.
In the absence of hormone, G proteins are
inactive and GDP (guanosine diphosphate)
is bound to α subunit. When a
peptide/protein hormone binds to a
receptor, the activated receptor induces a
conformational change in the G protein so
that GDP is released and GTP (guanosine
triphosphate) is now bound to the α
subunit. GTP binding to α subunit causes
dissociation of the α subunit from the
heterotrimeric complex; α subunit now
dissociates from the receptor and from
the βγ dimer, and moves along the
membrane. In the plane of the
membrane, the α subunit interacts with
downstream effectors such as adenylyl
cyclase and phospholipases
(phospholipase C and phospholipase A2).
• There are at least 16 Gα proteins that may
activate different types of effector
proteins.
(A) G proteins coupled to adenylyl cyclse
• A rather ubiquitous Gα protein is called
Gs-α, which stimulates the membrane
enzyme adenylyl cyclase. Adenylyl cyclase
then acts on the cellular ATP to convert it
into the cAMP; cAMP then acts as the
“second messenger”. The cyclic AMP
activates an enzyme, protein kinase A
(PKA or cAMP-dependent protein kinase).
PKA, in turn, catalyzes the
phosphorylation of various cellular
proteins, ion channels, and transcription
factors. This phosphorylation alters the
activity or function of the target proteins
and eventually leads to a desired cellular
response. E.g. parathyroid hormone.
In some instances, catalytic subunits
of PKA may also enter the nucleus,
where they phosphorylate and
activate the transcription factor,
cAMP response element-binding
protein (CREB protein). Phospho-
CREB then increases the
transcriptional rate of genes
encoding specific enzymes.
Synthesis of those specific enzymes
will increase.
• Some GPCRs couple to Gi-α, which inhibits
adenylyl cyclase. This decreases the levels
of cAMP. E.g. norepinephrine (NE) acting
via α2 receptors, dopamine acting via D2
receptors.
(B) G proteins coupled to phospholipase C
• A third major hormonal signaling pathway
involves receptors coupling to Gq-α,
which activates phospholipase C.
Phospholipase C acts on the phosphatidyl
inositol biphosphate (PIP2) and generates
two second messengers, the di-acyl
glycerol (DAG) and inositol triphosphate
(IP3). DAG stimulates protein kinase C
(PKC). IP3 binds to a receptor on the
endoplasmic reticulum membrane and
triggers the release of Ca++ from
intracellular stores; Ca++ activates
calmodulin-dependent protein kinases.
E.g. ADH, angiotensin II, TSH.
(C) G proteins coupled to phospholipase A2:
Some peptide hormones (e.g. TRH)
activate phospholipase A2. PLA2 then
cleaves membrane phospholipids to
produce lysophospholipid and arachidonic
acid. Arachidonic acid is converted, by
certain enzymes, into a variety of
biologically active eicosanoids (PGs,
prostacyclins, thromboxanes, and
leukotrienes.)
• G protein signaling is terminated by: (i)
intrinsic GTPase activity, (ii)
desensitization and endocytosis of the
receptor. GPCR kinases are the enzymes
that phosphorylate the intracellular
domain of the GPCRs. This recruits
proteins called β-arrestins. β-arrestin
binding inactivates the receptor, and also
the receptor is internalized into the cell by
endocytosis. This is an important
mechanism for hormonal desensitization
of a cell after exposure to excessive
hormone (decreased response of target
cells to hormone).
{Another type of G protein is a small,
monomeric G protein or low-molecular-
weight proteins. They play an important
role in many signaling pathways. They are
classified into 5 families: Ras, Rho, Rab,
Ran, and Arf.}
Second messenger systems:
Adenylyl cyclase-cyclic Phospholipase
AMP system system
ACTH Angiotensin II
TSH ADH
FSH & LH GnRH
PTH Oxytocin
Glucagon TRH
Calcitonin GH-RH
Secretin
catcholamines

[As already mentioned, norepinephrine

(via α2 receptors) and dopamine (via D2
receptors, produce their effects by
decreasing cAMP. Epinephrine (via β1 & β2
receptors) exerts its effects by increasing
cAMP. Angiotensin II, ADH, TRH exert
their effects via the second messengers
IP3 and DAG.]

2. CATALYTIC RECEPTORS:
Many hormones and growth factors
bind to cell membrane receptors that
themselves have enzymatic activity (or
they are a part of an enzyme complex)
on the cytoplasmic side of the
membrane. 5 classes of such catalytic
receptors have been identified:
a. Receptor guanylyl cyclase – When a
hormone combines with the receptor, the
receptor guanylyl cyclase catalyzes the
formation of cGMP (cyclic guanosine
monophosphate) from the intracellular
GTP. E.g. Receptor for atrial natriuretic
peptide (ANP). cGMP is used as a second
messenger by nitric oxide (NO) and
guanylin.
b. Receptor serine/threonine kinase – When
a ligand binds with the receptor, the
receptor phosphorylates serine and/or
threonine residues on cellular proteins.
E.g. receptors for the transforming
growth factor (TGF)-β family, which
includes the hormones anti-mullerian
hormone and inhibin.
c. Receptor tyrosine kinase – When a
hormone binds with the receptor, the
receptor phosphorylates tyrosine residues
on themselves and other proteins. E.g.
Receptors for insulin and insulin-like
growth factors (IGFs).
d. Tyrosine kinase-associated receptor –
When a hormone binds with the receptor,
the receptor interacts with cytoplasmic
(that is, non-membrane-bound) tyrosine
kinases. E.g. receptors for growth
hormone (GH), prolactin, erythropoietin,
and leptin.
e. Receptor tyrosine phosphatase – These
receptors cleave phosphate groups from
tyrosine groups of cellular proteins.

• SIGNALING FROM INTRACELLULAR

RECEPTORS -
- The steroid hormones, thyroid hormones,
and vitamin D act through intracellular
receptors.
- A steroid hormone enters the cytoplasm
of a cell, where it binds with a specific
receptor protein.
- The (receptor + hormone) complex then
diffuses into or is transported into the
nucleus; it binds to specific DNA
sequences in the nucleus. Thus, these
intracellular receptors act as
transcriptional regulators.
- There is activation of specific genes to
form mRNA.
- The mRNA diffuses into the cytoplasm
where it promotes the translation process
at the ribosomes to form new proteins;
these proteins then function as enzymes
or carrier proteins that in turn activate
other functions of the cells.
- The glucocorticoid receptor (GR) and
mineralocorticoid receptor (MR) are
mainly cytoplasmic; the estrogen and
progesterone receptors are primarily
nuclear. The thyroid hormone receptor
(THR) and retinoic acid receptors (RXR)
are bound to DNA in the nucleus. The
remarkable feature of nuclear receptors is
that they bind to specific DNA
sequences – called hormone response
elements (HREs) – in the regulatory region
of responsive genes.

Regulation of hormone sensitivity:

- A hormone may decrease the number or
affinity of receptors for itself or for
another hormone. This is “down-
regulation” of receptors.
- A hormone may increase the number or
affinity of receptors for itself or for
another hormone. This is “up-regulation”
of receptors.
Feedback regulation of the hormone
secretion:
- Hormone secretions are regulated
predominantly by “negative feedback”
loops. (Some exceptions - LH-estrogen
positive feedback before ovulation;
oxytocin release during parturition.)