Mechanism of action of hormones

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Hormones integrate coordinated response to stimulus
• Stimulus (change in environment)
• Hierarchical regulated hormone release
• Target cell: Hormone-receptor
• Intracellular signal
• 2nd messenger
• Tyrosine kinase
• Hormone-receptor complex
• Effect on target cell
• Protein level: modification, ion channels, translocation
• Gene level: induction, repression
• Coordinated response to the stimulus
homeostasis
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Features of hormone classes
Class I Class II
Type Steroids, iodothyronines, Peptides,
carcitriol, retinoids catecholamines
Solubility Lipophilic Hydrophilic
Transport protien Yes No
Plasma half-life Long (hours to days) Short (minutes)
Receptor Intracellular Plasma

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Signal transduction, class I
hormones
• Intracellular receptors
• Receptor (inactive or active)
• Hormone-receptor complex is signal
• Modulate gene expression
• Induce or repress gene transcription
• e.g., Glucocorticoids, Thyroid hormones, calcitriol, retinoids

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 Signal transduction,
•glucocorticoids
Cytoplasmic receptors
• Inactive receptor: heat shock protein (hsp) bound
• Hormone binding dissociates hsp, NLS exposure
• Receptor homodimerization
• Nuclear translocation
• Homodimer binds at HRE (GRE)
• high affinity
• TAD of dimer binds transactivator proteins
• Gene transcription
• Effects e.g., catabolic processes, hyperglycemic effects

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Signal transduction, thyroid hormone
• Nuclear receptors
• Regulated by coregulators (NCoR1 & SMRT)
• Thyroid receptor forms heterodimer with retinoid X receptor
• In absence of hormone:
• Dimer interacts with corepressors (NCoR1 or SMRT) & bind HRE (TRE)
• E.g., NCoR1 recruits histone deacetylase (compact chromatin)
• Gene repressed
• Hormone binding to dimer:
• Dissociation of corepressors
• Recruitment of coactivators
• Gene transcription
• Effects: e.g., increase
metabolic rate
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TR-RXR
activity

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 DNA sequences of
HREs

Effects of cAMP on gene transcription are mediated by the protein cyclic AMP response element binding protein (CREB).
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Specificity of
HREs
• GRE, PRE, MRE, and ARE: similar HRE
• Specificity
• Intracellular concentration of hormone or receptor
• Flanking DNA sequences
• Other accessory elements
• Thyroid hormones, retinoic acid, and vitamin D: similar HRE
• Specificity: spacing between repeats
• VDRE (N=3)
• TRE(N=4)
• RARE (N=5)

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Signal transduction, class II
hormones
• Cell surface transmembrane receptors
• Intracellular signal
• Second messengers
• Receptor tyrosine kinase
• Effects: protein level, gene level

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 G-protein-coupled receptors
•(GPCRs)
Receptor interacts with G-proteins
• G proteins:
• Heterotrimeric (αβ subunits)
• Inactive (GDP)
• Active (GTP)
• Inbuilt GTPase activity
• Second messengers (cAMP, DAG, IP3, cGMP & Ca2+)

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G proteins
• Relay signals form GPCRs to inside cells
• G protein families (Gs, Gi, Gq, and G12)
• Adenylate cyclase (ADCY) and phospholipase C (PLC)
• Gαs stimulates all isoforms of ADCYs
• Ten isoforms in humans (nine membrane-bound, one soluble)
• Gαi inhibit ADCY5 and ADCY6 isoforms
• Gq activates phospholipase C
• G12 role in diverse cellular processes such as platelet activation

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• .. q s

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G-protein coupled receptor signaling
• Hormone binds to the G protein-coupled receptor

• Receptor activation

• -GDP exchange for -GTP & dissociation from  subunit

• Activation of ADCY or phospholipase C

• 2nd messenger generation & effects (enzyme, ion channel, gene expression)

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G-protein coupled receptor
• 7 transmembrane helical domains

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Receptor activation

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Activation of
ADCYs
• E.g., TSH, glucagon and catecholamines

• Gs activates adenylate cyclase

• cAMP from ATP

• Allosteric activation of protein kinase A by cAMP

• Phosphorylation of effector proteins

• Cellular responses e.g., glycogenolysis, lipolysis 20

Receptor activation cAMP as second messenger

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 cAMP
•effects
Activation or inhibition of target proteins
• Phosphorylation of target proteins (Ser & Thr )
• Gene transcription:
• Mediated by cAMP response element binding protein (CREB)
• Nonphosphorylated CREB binds CRE
• weak transcription activator
• Phosphorylation of CREB by cAMP dependent PKA & coactivator
binding
• Potent transcription activator

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Termination of
signaling
• Phosphodiesterase hydrolyzes cAMP to AMP
• Methylxanthine (e.g., caffeine) prolong hormone actions, inhibit
phosphodiestarase

• Remove phosphate groups: phosphatases

• GTPase activity of -subunit (GTP to GDP)

• Cholera toxin inhibits GTPase activity
• Adneylate cyclase constantly activated

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cAMP
formation

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ADP-ribosylation by cholera
toxin

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IP3 and DAG as second
messengers

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 Activation of phospholipase
C activation
• GPCR
• Antidiuretic hormone
• 1- catecholamines
• Acetylcholine
• PLC activation Gq
• IP3 & DAG formation
• IP3 binds IP3R on ER
• Opening of Ca2+ channel & Ca2+
release into cytosol
• DAG activates PKC
 PLC
•activation
Vasopressin, acetycholine

Gq
+ • DAG activates protein kinase C

• IP3 binds IP3R (IP3-gated Ca2+ channel) on ER

• Channel opening & Ca2+ release to cytosol
• Ca2+ binds calmodulin
• Ca2+-calmodulin dependent protein kinases
• Phosphorylation
• E.g., phosphorylase kinase
Hormones & protein
Kinases
•PKA, PKC, and Ca -calmodulin-kinases
2+

• Phosphorylate serine and threonine residues on target proteins

• Role in hormone action

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cGMP as second messenger
• Natriuretic peptide (NP)
• Atrial Natriuretic peptide (ANP): highly express in cardiomyocytes
• Regulates salt-water balance and blood pressure
• Diuresis (renal sodium and water excretion)
• Vasodilation
• Inhibition of aldosterone secretion
• Brain natriuretic peptide (BNP):highly expressed in brain
• C-type Natriuretic peptide (CNP): highly expressed in brain
• Guanylate cyclase forms cGMP from GTP
• Soluble: role in NO signaling
• Membrane bound: activated by natriuretic peptides binding to receptors
• Potent vasodilators: Nitroprusside, nitroglycerin, nitric oxide, sodium
nitrite and sodium azide
• Increase cGMP
• Inhibitors of cGMP phosphodiesterase
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Natriuretic peptide receptors
• Three types (A, B, C)
• NPR-A: principal receptor of ANP & BNP
• synthesis of cGMP
• NPR-B: principal receptor of CNP
• NPR-C: ligand selectivity preference is ANP > CNP ⩾ BNP
• controls local natriuretic peptide concentrations via receptor-mediated
internalization and degradation

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Receptor Protein Tyrosine
Kinase
• Insulin, EGF, and IGF-I receptors

• Hormone-receptors interaction activates intracellular kinase activity

• Phosphorylation outside the catalytic domain (protein docking and

activation)

• Initiating of downstream signaling cascade

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 Insulin
•signaling
Insulin binding & activation of Receptor Tyrosine Kinase

• Autophosphorylation (Tyr on catalytic domain)

• IRS phosphorylation (Tyr) & protein docking

• Activation PI3K pathway and downstream signaling (Ser, Thr)

• mTOR is the main downstream target of PI3K pathway
• integrates nutrient and growth factor signals
• mTOR1: activated by nutrients such as amino acids & glucose
• Promotes anabolic processes
• mTOR2: activated by insulin, promotes cell survival and proliferation

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Insulin signaling pathways and effects 38
Receptor down regulation
• Prolonged exposure to a hormone decreases the cell’s response to
that level of hormone:

• Induce endocytosis and destruction of receptors in lysosomes

• Rapid inactivation of the receptors, e.g., receptor phosphorylation

• Production of inhibitor that blocks the transduction process

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Receptor upregulation
• Increase in the number of receptors when levels of the hormone is
low for some time.

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