Hormone Biochemistry, 

Synthesis & Release
Lecture 3
Lecture 3

(Serotonin)
 Lecture Outline
Lecture Outline
• Introduction
Introduction to hormone biochemistry
to hormone biochemistry
• Biosynthesis and secretion of polypeptide 
hormones
• Biosynthesis and secretion of steroid 
hormones
• Hormone transport mechanisms
 Endocrine Signaling
Endocrine Signaling

How are hormone 
synthesized in endocrine 
th i d i d i
cells and tissues?
 Hormone Biochemistry
• Not all hormones are the same!
Types of Hormones:
f

(T3))

(Serotonin)

Amino acid derivatives Polypeptide hormones

(Prostaglandin E1)
(Prostaglandin E1)
(Testosterone)
Fatty acid derivatives
Steroids (i.e., prostaglandins)
 Amino Acid Derivatives

• Hormones
Hormones such as catecholamines, 
such as catecholamines
indoleamines and thyroid hormones are 
formed by small modifications to amino acids
formed by small modifications to amino acids
• These hormones are small in molecular size
• Some are lipid soluble (thyroid hormones), 
S li id l bl ( h id h )
while others are lipid insoluble (serotonin, 
melatonin, epinephrine)
l i i hi )
 The Catecholamines: 
dopamine, epinephrine 
d i i hi
and norepinephrine

a catechol
(unsaturated six carbon ring)
(unsaturated six‐carbon ring)

Dopamine, epinephrine (adrenaline), 
and norepinephrine (noradrenaline) 
are derived from the amino acid 
tyrosine and are released into 
circulation to have endocrine 
functions
 Indolamines are derived from the 
The Indolamines:  amino acid tryptophan
serotonin (5‐HT) and 
t i (5 HT) d
melatonin

MAO enzyme
 The Indolamines: 
serotonin (5‐HT) and 
t i (5 HT) d
melatonin

Monoamine oxidase
(MAO) enzymes are 
key in the conversion 
of serotonin (5‐HT)
of serotonin (5 HT) to 
to
the inactive 5‐HIAA.  
This is why MAO 
inhibitors (MAOIs) are
inhibitors (MAOIs) are 
used as 
antidepressant drugs
 tyrosine Thyroid Hormone 
Synthesis
h
Begins with the amino acid tyrosine

3‐monoiodotyrosine (MIT) 3,5‐diiodotyrosine (DIT)

triiodothyronine (T3)

thyroxine (T4)

reverse T3 (rT3)
 Amino Acid Derivatives ‐ Summary

• Includes: 
– catecholamines (epinephrine, norepinephrine)
– indoleamines (serotonin, melatonin)
– thyroid hormones (T4, T3)
th roid hormones (T4 T3)
• These hormones are all formed by modifications to 
the amino acids tyrosine
the amino acids tyrosine
– Tyrosine (catecholamines, thyroid hormones)
– Tryptophan (indolamines)
Tryptophan (indolamines)
• Some are lipid soluble (thyroid hormones), while 
others are lipid insoluble (serotonin, melatonin, 
p (
epinephrine)
 Hormone Biochemistry
• Not all hormones are the same!
Types of Hormones:
f

(triiodothyronine)

(Serotonin)

Amino acid derivatives Polypeptide hormones

(Prostaglandin E1)
(Prostaglandin E1)
(Testosterone)
Fatty acid derivatives
Steroids (i.e., prostaglandins)
An example of a polypeptide hormone:
 Protein Hormone Synthesis
• Transcription – RNA synthesis 
f
from DNA
DNA
– Post‐transcriptional modification: 
controlled modification of RNA to
controlled modification of RNA to 
form mRNA from excision and 
splicing

• Translation – assembly of amino acids via specific base 
pairing of the anticodons
p g of the carrier amino acylated
y
transfer RNAs to the correspond codons of the mRNA 
(creation of the polypeptide chain)
RNA polymerase 
(RNAP) Gene Transcription (DNA     RNA)

Step 1

Step 2

Step 3
Gene Translation (mRNA      Polypeptide Protein)
 5´ UTR  Hormone coding region (uppercase)
(lowercase) (signal peptide underlined)

-81 -57
atcctgaaaccctgccagctataaATGTCTCCTCTGTATGTGGTTGGCATGCTGGGACTTTTGATG
M S P L Y V V G M L G L L M
• Coding and 5´
and 3´
+1
AAGATATCAGCACCTATGTGTGCCCCCACTGAGTCCACCATTTACTTCGAGAGACAG
K I S A P M C A P T E S T I Y F E R Q

GAGTGTAACTACTGTGTGGCTGTCAACACCACCATCTGCATGGGCTTCTGCTTCTCC
E C N Y C V A V N T T I C M G F C F S
untranslated 
AGGGACAGTAATGTGAAGGAGTTGGTGGGTCCTCGTTTCCTGGTTCAGAGAGGCTGC
R D S N V K E L V G P R F L V Q R G C
regions (UTRs) 
ACCTATCAGGAAGTCGAGTATCGGACGGCCATCTTGCCTGGCTGTCCTTCATATGCA
T Y Q E V E Y R T A I L P G C P S Y A
shown in the 
GATCCTCACTTCACCTATCCAGTGGCACTTAGTTGCCACTGCAGCACCTGTAACACC
D P H F T Y P V A L S C H C S T C N T
cDNA for 
CACAGTGACGAATGTGCCCACAAAACCAGCAGCGCTGCAAGGAAATGCTCCAAACCT thyrotropin from
thyrotropin from 
H S D E C A H K T S S A A R K C S K P

GTCCGACATCTGTACCCCGACCCCGAGGAGAACAGTTACATCCAGCCATATTGGGAA fathead minnow
V R H L Y P D P E E N S Y I Q P Y W E

CAGTACGAGTAAtgtcactttttgggaaatgggctcattttacaatttttgccgtttttgaatcaattcggccgattttctgatat
CAGTACGAGTAAt t ttttt t t tttt ttttt ttttt t tt tttt t t t
Q Y E stop

ttggagtagtgtcggtagccagaccgtttctggtctcattgacttccatagtagggaaaaatatatactatggaagtcaatgagac
cagaggctgtctggttgcagatattcttctgaatatcaggaagtcggctgaatggattcgaaaacggtaaaactctgtttaactct
gggggagttgtaaaatgagcctatttccaaaaaagtggagtatccctttaatttaaaacaaagtgtgtgtgtgttgtagcagttga
ctgggtgcaattgtttgttagataatcattgagtaacctttcagttacctaacttattatggttgctgtatttatatcaagcatctgatgtc
ttttatgaggccatagtggttaaaaaatggtgtggttcaatacttaagcttttatggataatagaccttctgccaacttaatgacgctt
3 UTR 
3´ UTR
aacgttggatcggcagtttttctataaaaaataaaagtaaaatccttc–(Poly A27) (lowercase)

Lema et al (2008) Gen. Comp. Endocrinol.

 Types of Peptide Hormone mRNAs
Types of Peptide Hormone mRNAs
• Most polypeptide hormones are transcribed as a 
preprohormone
h mRNA that has a signal peptide 
RNA th t h i l tid
attached.  This signal peptide is cleaved as the protein 
hormone is translated and translocated across the rough  g
endoplasmic reticulum (RER) membrane.

• Many
Many polypeptide hormones are translated
polypeptide hormones are translated as a as a
prohormone protein that are precursors to the active 
molecule.  The prohormone is converted into the active 
hormone within the cell where it is produced, in the 
blood,  or in target tissues.  Note that many 
prohormones possess some of the activity of the mature
prohormones possess some of the activity of the mature 
hormone. 
 Preprohormones:
conversion to active hormones
i i h
Examples of prohormones:
Functions of a prohormone:
1) Further proteolytic processing in some cells may produce 
other biologically active peptides – type of processing 
other biologically active peptides  type of processing
depends on cell type
2) May facilitate transport and sorting in cell
3) May facilitate folding of the molecule
Prohormone postranslational processing can be 
complex but is key to hormone function!
complex, but is key to hormone function!
Alternate Splicing of Genes
during mRNA synthesis
during mRNA synthesis
• Genes are spliced to remove non‐
coding introns immediately following 
coding introns immediately following
transcription 

• Many hormone‐associated genes 
show alternate splicing: the 
creation of multiple mRNA variants
creation of multiple mRNA variants 
for the same gene. 

• Some of these mRNA slice 
Some of these mRNA slice
variants are translated, created 
slight variations in the proteins 
(especially common with hormone 
receptor proteins)
thyrotropin β‐subunit from goldfish
(Yoshiura et al 1999 Fish Physiol Biochem 21, 201‐210)
Polypeptide Hormone Release
Polypeptide Hormone Release
 Characteristics of polypeptide hormones:
Characteristics of polypeptide hormones:

1) Postranslational
Postranslational processing can lead to cell and 
processing can lead to cell and
tissue specific expression
2)) Alternative splicing may contribute to different 
p g y
hormone isoforms (certainly common with 
hormone receptors)
3) Evolutionary divergence in polypeptide hormones is 
comparatively rapid
‐ Considerable taxonomic variation in types of polypeptide 
hormones – even when they have similar functions
 Polypeptide Hormones Show High Variation in Amino 
Acid Structure among Taxa

→
 Characteristics of Polypeptide Hormones
Characteristics of Polypeptide Hormones
• Polar, lipid insoluble
– Cannot pass through the lipid cell membrane
• Bind to receptor proteins on the cell surface
Bind to receptor proteins on the cell surface
– Binding of hormone to receptor activates 
membrane‐bound
membrane bound enzymes 
enzymes
 Hormone Biochemistry
• Not all hormones are the same!
Types of Hormones:
f

(triiodothyronine)

(Serotonin)

Amino acid derivatives Polypeptide hormones

(Prostaglandin E1)
(Prostaglandin E1)
(Testosterone)
Fatty acid derivatives
Steroids (i.e., prostaglandins)
Steroid Hormone Synthesis
Pathways of Steroid Hormone Synthesis
(See Handout)
Na+/water
stress responses
p
homeostasis

sex differences
in phenotype
 Steroid Synthesis 
y
Summary ‐ Handout
Steroid Hormone
Synthesis

Take-home point 1:

Steroid synthesis begins with

the conversion of cholesterol
to pregnenolone
 Transport of cholesterol from cytoplasm 
across mitochondrial membrane is the 
rate‐limiting step in steroidogenesis

• Steroid
Steroid acute regulatory 
acute regulatory
protein (StAR) – a transport  StAR
protein that carries the 
lipophilic cholesterol from 
the cytoplasm into the 
mitochondria
– StAR can be up‐ and down‐
regulated by hormones and 
environmental factors
environmental factors
Steroid Hormone
Synthesis

Take-home point 2:

Corticosteroids, androgens
g and
estrogens are all generated
from the same precursor
(pregnenolone) through
conversion enzymes
• Tissue and cell variation in expression of conversion 
enzymes determines tissue‐ and cell‐specific 
steroidogenesis

• Regulation of 
steroidogenesis
can occur by up‐ or 
down‐regulating 
conversion 
conversion
enzymes
Differences in 
steroidogenesis
pathways b/w gonads 
and adrenal cortex 
Steroid Hormone Synthesis
Synthetic Steroids
Synthetic Steroids
 Characteristics of Steroid Hormones 
(and Thyroid Hormones)

• Non‐polar, lipid soluble
– pass directly through the lipid cell membranes
pass directly through the lipid cell membranes
• Usually bind to protein receptors in the 
cytoplasm or nucleus
cytoplasm or nucleus
– recent evidence for membrane steroid receptors 
as well
as well
 Hormone Biochemistry
• Not all hormones are the same!
Types of Hormones:
f

(triiodothyronine)

(Serotonin)

Amino acid derivatives Polypeptide hormones

(Prostaglandin E1)
(Prostaglandin E1)
(Testosterone)
Fatty acid derivatives
Steroids (i.e., prostaglandins)
 Essential Fatty Acid Substrate Derivate 
H
Hormones (Eicosanoids)
(Ei id )

Prostaglandin E1 Thrombaxane A2
(regulate vasotconstriction/dilation; (regulate inflammation)
hypertension)

Leukotriene B4 Juvenile Hormone 3

(regulate inflammation) (regulates insect development )
• There are over 20 pathways for conversion of essential fatty acids to hormones
• Fatty acids compete for enzyme availability
• Omega-3
g fattyy acid derivatives lessen inflammation caused by
y omega-6
g fattyy
acid derivates
• Low dietary intake of less inflammatory essential 
fatty acids such as Omega 3 has been linked to
fatty acids such as Omega‐3 has been linked to 
inflammation‐related diseases and perhaps even 
mental illnesses.
• Increased dietary Omega‐3 helps prevent 
cardiovascular disease, hypertension, rheumatoid 
arthritis and can ease symptoms in psychiatric 
disorders 
 Lecture Summary
Lecture Summary
• 4 types of hormones:
– Amino acid derivatives
– Polypeptide hormones – coded by genes
– Steroid hormones cholesterol derived
Steroid hormones ‐ cholesterol deri ed
– Lipid derived

• These hormone differ in mechanisms for synthesis 
and regulation
• The biochemical properties (polarity, lipophilicity) 
vary widely
– Implications for transport and mode of action on cells
 Lecture Questions
1) What are the major classes of hormones, and
how do they differ?
2) How are polypeptide hormones synthesized?

3) How are steroid hormones synthesized? What

regulates
g tissue-specific
p synthesis
y of steroids?
4) Why is there so much variation among taxa in
the structure of polypeptide hormones
hormones, but not
among steroid hormones or amino acid
derivative hormones?