Endocrine System Diversity

Peter Bryan E. Dela Cruz, MD, FPCP, CSPSH, DPCOM

Section of Biochemistry
College of Medicine
Central Philippine University
 Outline
• Introduction
• Diversity of Endocrine System
• Synthesis of Hormones
- from cholesterol, amino acid, peptides
• Storage and Secretion
• Hormone Function
• The survival of multicellular organisms depends on their ability to
adapt to a constantly changing environment.

• Intercellular communication mechanisms are necessary

requirements for this adaptation.

• The nervous system and the endocrine system provide this

intercellular, organism-wide communication.

• The nervous system - originally viewed as providing a fixed

communication system

• The endocrine system -supplied hormones, which are

mobile messages. For example, neural regulation of the endocrine
system is important in the production and secretion of some
hormones; many neurotransmitters resemble hormones in their
synthesis, transport, and mechanism of action; and many
hormones are synthesized in the nervous system.
 ABBREVIATIONS
• ACTH Adrenocorticotropic hormone
• ANF Atrial natriuretic factor
• cAMP Cyclic adenosine monophosphate
• CBG Corticosteroid-binding globulin
• CG Chorionic gonadotropin
• cGMP Cyclic guanosine monophosphate
• CLIP Corticotropin-like intermediate
lobepeptide
• DBH Dopamine -hydroxylase
• DHEA Dehydroepiandrosterone
• DHT Dihydrotestosterone
• DIT Diiodotyrosine
• DOC Deoxycorticosterone
• EGF Epidermal growth factor
• FSH Follicle-stimulating hormone
• GH Growth hormone
• IGF-I Insulin-like growth factor-I
• LH Luteotropic hormone
• LPH Lipotropin
• MIT Monoiodotyrosine
• MSH Melanocyte-stimulating hormone
• OHSD Hydroxysteroid dehydrogenase
• PNMT Phenylethanolamine-N -
methyltransferase
• POMC Pro-opiomelanocortin
• SHBG Sex hormone-binding globulin
• StAR Steroidogenic acute regulatory
(protein)
• TBG Thyroxine-binding globulin
• TEBG Testosterone-estrogen-binding
globulin
• TRH Thyrotropin-releasing hormone
• TSH Thyrotropin-stimulating hormone
Hormones
- Greek term meaning “ to set to motion”
- elicit cellular response and regulate
physiologic processes

Endocrine System
- no anatomic lines
- uses mobile messages through hormones
• Classic Endocrine Organs
Hypothalamus
Pituitary
Thyroid
Parathyroid
Pancreas
Adrenals
Gonads

• Others
Small Intestines
Stomach
Kidneys
Heart
Placenta
Adipose Tissue
 Diversity of
Endocrine System
Classification of Hormones
- according to chemical composition, solubility,
location of receptors and nature of signal used
 Site of Hormone Synthesis

• Organs
1. discrete organs – pituitary, thyroid,
adrenals
2. organs that perform 2 closely related
functions – ovary, testes
• Specialized cells
- Ex. C cells of the thyroid - calcitonin
• Parenchymal cells of more than 1 organ
- Ex. Calcitriol – skin, liver, kidney
 Chemical Diversity

• Cholesterol derivatives – cortisol,

androgens, calcitriol
• Amino acid derivatives – dopamine,
epinephrine, thyroid hormones
• Peptides – insulin, PTH, growth hormone,
ACTH
• Glycoproteins – FSH, TSH,LH, HCG
- αβ heterodimer
- β chain impart hormone uniqueness
 Synthesis and Modification

• Synthesized in the final form and secreted

immediately
• Synthesized in final form and stored in
producing cells
• Synthesized from precursor molecules –
processed and secreted upon physiologic cue
• Converted to active form precursor in the
periphery
Steroid Hormones
 Adrenal Steroidogenesis
3 Classes of Steroids
1. Mineralocorticoids
2. Glucocorticoids
3. Androgens
- synthesized from cholesterol
- uptake of cholesterol mediated by LDL receptors
- cholesterol stored in lipid droplets of the
adrenals
Note:
Upon ACTH stimulation esterase is stimulated
and free cholesterol is transported to the
mitochondria.
 All mammalian steroid hormones are formed from
cholesterol via pregnenolone.
Mineralocorticoid Synthesis
Glucocorticoid Synthesis

17 α hydroxylase

3β OHSD isomerase

21 hydroxylase

11β hydroxylase
Androgen Synthesis
 Testosterone Synthesis
DHEA Pathway

17, 20 lyase

17 β OH dehydrogenase,
Δ 5,4 isomerase
Androgen Synthesis
Testicular Steroidogenesis (Progesterone Pathway)

Pregnenolone

Progesterone

17α Hydroxyprogesterone

Androstenedione

Testosterone

DHEA Pathway appears to be the most useful in human testes.

Testosterone Metabolism

Aromatase

5α reductase
Ovarian Steroidogenesis

Estriol
Calcitriol Synthesis
Tyrosine Derivatives
 Catecholamines
• 3 Amines:
1. dopamine
2. norepinephrine – present in organs innervated by
sympathetic nerves
3. epinephrine – major product of adrenal medulla
- 80% of catecholamines of the adrenal
medulla

• Steps in Epinephrine Synthesis

1. Ring hydroxylation
2. Decarboxylation
3. Side chain hydroxylation to form NE
4. N-methylation to form epinephrine
Catecholamine Synthesis
Catecholamine Catabolsim
• Catechol-O-methyltransferase (COMT)
- addition of methyl group at 3 position of benzene ring
- Metabolites: homovanillic acid (dopamine)
normetanephrine (norepinephrine)
metanephrine (epinephrine)
• Monoamine oxidase (MAO)
- oxireductase
MAO A – deaminates serotonin, epinephrine and norepinephrine
MAO B – active against phenylamine and benzylamine
Important Catecholamine Metabolites
1. Metanephrines – methoxyderivatives of E and NE
2. Vanillyl mandelic acid (VMA) – O methylated deaminated
product of E and NE
 Thyroid Hormone Synthesis
A. Iodine Uptake
- first step in thyroid hormone synthesis
- thyroid able to concentrate I- against a strong
gradient
- linked to Na+, K+ ATPase dependent I- transporter
Note: transporter mechanism inhibited by percholate,
perrhenate, pertechnetate and thiocyanate

B. Oxidation of I-
- oxidized to a higher valence state
- involves thyroperoxidase
- occurs in the luminal surface of the follicular cell
- inhibited by thiourea drugs
C. Organification of I2
Thyroglobulin – glycosylated protein precursor of T3
and T4
- 2 subunits each with potential site of iodination
Thyroglobulin + I2 → MIT or DIT
D. Coupling of Iodotyrosyls
MIT + DIT →T3
DIT + DIT → T4

Note: In the periphery, T4→ T3

Peptide Hormones
Insulin

• Polypeptide consisting of
2 chains linked by 2
interchain disulfide
bridges

• 3rd interchain disulfide

bridge connects residues
6 and 11 in chain A

• Synthesized as a
prohormone
 Parathyroid Hormone

PreproPTH
↓ → 25 amino acid peptide
ProPTH
↓ → pro extension
Mature PTH
storage ← ↓ → degraded
Secreted

Note: Rate of degradation of PTH decreases with

decrease serum calcium.
Angiotensin II
 POMC Protein

• Expressed in the anterior and intermediate lobes of the

pituitary
• Synthesized as 285 amino acid precursor
• Cleavage produces peptides that act as hormones,
neurotransmitters or neuromodulators
• Processed differently in different parts of the pituitary

Note :
Diversity of products is due to many dibasic amino acid clusters
that are potential cleavage sites.

Each peptide is preceded by Lys-Arg, Arg-Lys, Arg-Arg or Lys-

Lys residues
Storage , Secretion
and Transport
 Storage and Secretion
1. Steroids – secreted as they are made
- no storage
2. Catecholamines – stored in granules in chromaffin
cells
- supply good for hours
3. PTH – stored in vesicles
4. Insulin – stored in pancreatic β cells
- supply good for days
5. Thyroid hormone – stored in colloid of follicular cells
- supply good up to 2 weeks
 Transport Proteins

Hormones
• Class I – hydrophobic
- have specialized plasma transport proteins
• Class II – hydrophilic
- do not require transport proteins

Bound hormones – provide circulating reservoir

- prolong half life
- restrict hormone access to certain sites
- inactive
• Thyroid
- most circulate bound to TBG
- TBG binds T4 with greater affinity
• Glucocorticoids
- Transcortin/CBG – main plasma binding protein
- increased by estrogens
- Deoxycorticosterone, corticosterone and
progesterone also bind with CBG
Note: Aldosterone has no specific transport protein.
• Gonadal Steroids
- SHBG – increased by estrogens, hyperthyroidism
and certain liver diseases
- decreased by androgens, advancing age and
hypothyroidism
Note:
Testosterone binds with SHBG with greater affinity
than estradiol.
Estrogens bind to SHBG.
Progestins bind to CBG.
Hormone Function
 Three General Areas

• Growth and Differentiation

- multiple hormones and nutritional factors
• Maintenance of Homeostasis
1. Thyroid hormone – controls 25% of basal metabolism
2. Cortisol – exerts permissive action + its own effect
3. PTH – regulate calcium and phosphorus levels
4. Vasopressin – regulates osmolality
5. Mineralocorticoids – vascular volume & serum electrolytes
6. Insulin – maintains euglycemia
• Reproduction
 Stages of Reproduction
1. sex determination and fetal development
2. sexual maturation
3. conception, pregnancy, lactation
4. cessation of reproductive capability
 Hormone Actions
Peter Bryan E. Dela Cruz, MD, FPCP, CSPSH, DPCOM
Section of Biochemistry
College of Medicine
Central Philippine University
Outline

• Target Cell Concept

• Hormone Receptors
• Signal Generation
- Cell Membrane Receptors
- Nuclear Receptors
• Hormones and Transcription
• Termination of Hormone Action

55
Summary
• Hormones are synthesized from a number or precursor
molecules

• Hormones use a variety of signaling mechanisms to facilitate

adaptive responses.

• The hormone-receptor complex serves as initial signal for

members of the nuclear receptor family.

• Hormones that bind to cell membrane receptors generate a

variety of intracellular signals.

• Many hormone responses are accomplishes through

alterations in the rate of transcription of specific genes.
Target Cell Concept

57
Hormone Concentration
• Rate of synthesis and secretion
• Proximity of target cell
• Dissociation constants with transport proteins
• Rate of clearance
Target Cell
• Number, relative activity and state of occupancy
of receptor
• Metabolism of the hormone in the cell
• Presence of other factors
• Up or down regulation of receptor
• Post receptor desensitization
58
Hormone Receptors

- Receptors discriminate precisely.

+ A target cell is defined by its ability to selectively
bind a given hormone to its cognate receptor.

Hormone-Receptor Interactions*:
1. Binding should be specific
2. Binding should be saturable
3. Binding should occur within the concentration
range of the expected biologic response

59
• Receptors have at least 2 functional domains:
1. Recognition domain
2. Coupling domain

• Signal transduction occurs in 2 ways:

1. Group I hormones
- intracellular receptors
- ligand receptor complex provides the signal
2. Group II hormones
- plasma membrane receptors
- generate a signal that regulates intracellular
functions

60
• Functional Domains of Steroid & Thyroid
Receptor
1. Binds the hormone
2. Binds specific DNA regions
3. Interaction with other coregulator proteins
4. Binding to one or more proteins that influence
the intracellular trafficking of the receptor

It is the coupling & hormone binding to signal

transduction that provides the first step in
hormone response.

62
Membrane Receptors

Major Groups:
1. Transmembrane GPCRs
2. Tyrosine kinase receptors
3. Cytokine receptors
4. Serine kinase receptors

63
Transmembrane GPCR
- binds to LH, PTH, TRH,
somastotatin,
catecholamines
1. Extracellular domain
- major binding site
2. Transmembrane
- hydrophobic helical
domains
- traverse lipid bilayer
3. Intracellular domain
- docking site for G
proteins

64
G Proteins

• composed of α, β, γ subunits
• α subunits contain guanine
nucleotide binding site
• Absence of hormone:
- G proteins complex is
inactive GDP bound form
• With hormone:
- GDP is replaced by GTP
- α dissociates and binds to
and activates effector

66
67
Tyrosine Kinase Receptors

• Insulin and other growth factors

• Undergoes autophoshorylation

68
Cytokine Receptors

PI 3K
MAP kinase pathway
Phospholipase C

69
Serine Kinase Receptors

• have features similar to tyrosine kinase

receptors
- N terminal extracellular domain
- single transmembrane domain
- C terminal intracellular domain possess
protein kinase activity
* They differ in enzymatic specificity.
• mediates activins, TGF β, mullerian inhibiting
substance and bone morphogenic proteins
• signals through proteins termed as SMADS

71
Nuclear Receptors

Receptors: Steroid RXR Partnered

Binding: homodimers heterodimers

Ligand: steroids Thyroid, Vit D,

retinoid
DNA element: Inverted repeat Direct repeats

Note: Carboxyl terminal hormone binding domain mediates

transcriptional control.
72
In RXR Partnered Receptors:
- corepressor proteins bind to the receptor in
the absence of hormone
- Hormone binding cause release of
corepressor and recruitment of activators

Receptor-Coactivator Complex
1. Recruit enzymes that modify chromatin
structure
2. Interact with transcription factors
3. Interact with transcription apparatus

73
Signal Transduction
Receptor → Transducer → Effector

+ Recognition of stimulus is the first step in the adaptive

response.
Stimulus
↓
↓ ↓
Group I Group II
↓ ↓
Hormone-receptor complex Many different
signals
↓ ↓
COORDINATED RES PONSE
Signal Generation

75
Group I Hormones
• Hormones diffuse through plasma membrane and
encounter intracellular receptors in target cells

• Uses ligand-receptor complex

• Hormone-Receptor complex undergoes activation

reaction

• Receptors are either in the cytoplasm or in the

nucleus

76
 Accelerated gene
transcription
HRE

77
 R

R
• DNA bound receptor is bound to corepressors
and actively represses gene transcription
• The association of ligand results in the
dissociation of the corepressors

79
Group II Hormones

• Receptor is located in the plasma membrane

• Use intracellular messengers
• 2nd Messengers
- cAMP
- cGMP
- Ca2+
- phosphatidylinositides
G Protein Coupled Receptors (GPCR)
cAMP
1. Adenylyl cyclase
- different peptide hormones either stimulate
or inhibit production of cAMP from adenylyl
cyclase
- 2 parallel systems converge upon a single
catalytic molecule (represented by C)
- each consists of a receptor and a regulatory
complex
 Hs Hi

Rs Ri

β β β β
αi
αs γ γ γ γ

αs ---GTP αi ---GTP

membrane
+ --
C

ATP cAMP cytoplasm

2. Protein kinase
- protein kinase A (PKA)
With cAMP binding:
4cAMP + R2C2 ↔ R2 . (4cAMP) + 2C

- R2C2 complex has no enzymatic activity

- active C catalyzes transfer of γ phosphate of
ATP to serine and threonine residues
 C
R

4cAMP
R
C

+ 2 C

ATP

protein phosphoprotein

phosphatase

Physiologic effect
3. Phosphoproteins
- effect of cAMP mediated phosphorylation-
dephosphorylation
- define response within a given cell
4. Phosphodiesterase
- terminate hormone action by hydrolysis of
cAMP to 5’-AMP
- ensures rapid turnover of cAMP
- inhibitors increase cAMP and mimic or
prolong actions of hormones
5. Phosphoprotein phosphatase
- inactivates phosphoprotein, terminating
hormone effect
cGMP

• Made from GTP by guanylyl cyclase

• Exist in soluble and membrane bound forms
• Used by atriopeptins
Guanylyl cyclase activated
↓
Increase cGMP
↓
activated
↓
Phosphorylation of smooth muscle proteins
Calcium or Phosphatidylinositol

Calcium metabolism
- Intracellular calcium concentration is low
- Ca2+ is restrained from entering the cell
- Mechanisms of intracellular concentration
control:
Na+/Ca2+ exchange
Ca2+ /proton ATPase pump
Note: Ca2+ /proton ATPase pump calcium from
the cytosol to the ER
Ways to Modify Cytosolic Ca2+
1. Hormones binding to receptors enhance
membrane permeability
2. Hormones indirectly promote Ca2+ influx by
modulating membrane potential
3. Ca2+ mobilized from the ER
Note: Discovery of calmodulin provided basis for
understanding how calcium interacts with cells.
Calmodulin
- calcium dependent
regulatory molecule
- 4 Ca2+ binding sites
- Full occupancy of the
binding sites leads to
conformational changes
and activation of
enzymes and ion
channels
- Involved in regulating
kinases and cyclic
nucleotide generation
and degradation
Calcium as mediator of Hormone Action
- effect of hormones blunted with decreased
intracellular calcium
- hormone effects can be mimicked by agents
that increase cytosolic calcium
- hormones influence cellular calcium flux
Ca/Calmodulin
Complex

Kinase
activation
Protein Kinase Cascade

- ligands involved in growth control,

differentiation and inflammatory response
- receptors with tyrosine kinase activity or
associated with proteins that are tyrosine
kinases
- preferentially phosphorylate tyrosine residues
- ligand-receptor complex initiates a cascade
- involve several protein kinases, phosphatases
and other regulatory proteins
Insulin
JAK/STAT Pathway(Cytokine Receptors)

• Used by growth hormone, prolactin,

erythropoietin, and cytokines

• Initiate action by activating tyrosine kinase

(activity not integral part of the hormone
receptor)

• Activate cytoplasmic tyrosine kinases

PI 3K
MAP kinase pathway
Phospholipase C
Serine Kinase Pathway

99
Hormones and Transcription
Observations:

1. Actively transcribed genes are in regions of open

chromatin.
2. Genes have regulatory regions where transcription
factors bind.
3. Hormone-receptor complex can be a transcription factor.
4. Other hormones generate signals that modify location,
amount and activity of transcription factor.
5. Nuclear reeceptor family act analogous to hormone
receptors.
6. Nuclear receptors interact with regulatory proteins.
• HRE (Hormone Response Elements)
- within a few hundred nucleotides upstream
of initiation site
- may be located within coding region
- associate with DNA elements and binding
proteins to function optimally
• Nuclear Receptor Proteins
- diverse set of transcription factors
- include hormone receptors
• Structural Features of Nuclear Receptor Proteins
1. centrally located DNA binding domain (DBD)
2. Multifunctional ligand binding domain (LBD)
3. Highly variable hinge region
4. amino acid termial region that contains AF1

Note: Nuclear receptor coregulators participate in

regulating transcription.
Ex. CPB, p160 proteins
Termination of Hormone Action
• Receptor mediated endocytosis
- Ex. Insulin receptors are internalized: some
are recycled but some are degraded
• Protein tyrosine phosphatases
- remove phosphate
- antagonize action of tyrosine kinase
• Serine/threonine kinases
- appears to inhibit action of tyrosine kinase

103
Summary
• Hormones are synthesized from a number or precursor
molecules

• Hormones use a variety of signaling mechanisms to facilitate

adaptive responses.

• The hormone-receptor complex serves as initial signal for

members of the nuclear receptor family.

• Hormones that bind to cell membrane receptors generate a

variety of intracellular signals.

• Many hormone responses are accomplishes through

alterations in the rate of transcription of specific genes.