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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)
• 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
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)
• 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?