Lecture 2

Mechanisms of Hormone Action

 Receptors: intracellular and membrane
bound receptors.
 Signal transduction
 Second messenger role in signal
transduction: cAMP, cGMP, lipids,
Calcium ions
 PART A

Receptors
Structure and Function
 The role of the receptor
• Globular proteins

• Located mostly in the cell membrane

• Receive messages from chemical messengers coming from other

cells (CNS)

• Transmit a message into the cell leading to a cellular effect

• Different receptors specific for different chemical messengers

• Each cell has a range of receptors in the cell membrane making it

responsive to different chemical messengers
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 The Role of the receptor…

Nerve Nerve
Signal

Messenger
Receptor

Response
Nucleus Cell
Cell

4
 The Role of the receptor…
• Neurotransmitters: Chemicals released from nerve
endings which travel across a nerve synapse to bind
with receptors on target cells, such as muscle cells
or another nerve. Usually short lived and
responsible for messages between individual cells

• Hormones: Chemicals released from cells or glands

and which travel some distance to bind with
receptors on target cells throughout the body

Note: Chemical messengers ‘switch on’ receptors

without undergoing a reaction
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 The role of the receptor….
• Receptors contain a binding site (cleft on the receptor
surface) that is recognised by the chemical messenger

• Binding of the messenger involves intermolecular bonds

• Binding results in an induced fit of the receptor protein

• Change in receptor shape results in a ‘domino’ effect

• Domino effect is known as signal transduction, leading to

a chemical signal being received inside the cell

• Chemical messenger does not enter the cell. It departs the

receptor unchanged and is not permanently bound 6
 The Binding Site
• A hydrophobic hollow or cleft on the receptor surface -
equivalent to the active site of an enzyme

• Accepts and binds a chemical messenger

• Contains amino acids which bind the messenger

• No reaction or catalysis takes place

Binding site

ENZYME

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 The Binding Site

Messenger
M

Induced fit

• Binding site is nearly the correct shape for the

messenger
• Binding alters the shape of the receptor (induced fit)
• Altered receptor shape leads to further effects - signal
transduction
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How does the Binding Site Change Shape?

Phe
Phe

H
O
H
O Ser
Ser CO2
CO2 Induced
Asp Fit Asp

 Before –
 Intermolecular bonds not optimum length for maximum binding strength
 After –
 Intermolecular bond lengths optimised

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 Induced Fit
• Binding interactions must be strong enough to hold the
messenger sufficiently long for signal transduction to
take place
• Interactions must be weak enough to allow the
messenger to depart
• Implies a fine balance
• Designing molecules with stronger binding interactions
results in drugs that block the binding site - antagonists
M M

RE RE
R

Signal transduction 10
 Main Types of Receptors

• Ion channel receptors

• G-protein-coupled receptors

• Kinase-linked receptors

• Intracellular receptors

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 1. Ion Channel Receptors
• Receptor protein is part of an ion channel protein complex

• Receptor binds a messenger leading to an induced fit

• Ion channel is opened or closed

• Ion channels are specific for specific ions (Na+, Ca2+, Cl-, K+)

• Ions flow across cell membrane down concentration

gradient

• Polarises or depolarises nerve membranes

• Activates or deactivates enzyme catalysed rxns within cell

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 Ion Channel Receptors…
Hydrophilic
tunnel

Cell
membrane

MESSENGER

RECEPTOR
BINDING
ION
SITE
CHANNEL
(open)
MESSENGER

Induced fit
Lock and opening
Ion Gate Ion of ion channel Cell Ion Ion Cell
Cell Cell
membrane channel channel membrane membrane channel channel membrane

Cell
Cell
 Ion Channel Receptors…

Transmembrane Proteins
Protein
TM4
subunits
TM1 TM3

TM3 TM2 TM1

TM4 TM2 TM2 TM4

TM1 TM3

TM3 TM2 TM2 TM1

TM4 TM1 TM3 TM4

TM2 of each protein subunit ‘lines’ the central pore

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 Gating

Binding site
Receptor Messenger

Induced
Cell
membrane fit Cell
membrane
‘Gating’
(ion channel
opens)
Five glycoprotein subunits
traversing cell membrane

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 Gating…
• Chemical messenger binds to receptor binding site
• Induced fit results in further conformational
changes
• TM2 segments rotate to open central pore

TM2 TM2
Cell
membrane

TM2
TM2

TM2
TM2
TM2 TM2
Transverse view Transverse view
TM2 TM2 TM2

TM2

Closed
Open
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 Gating…
• Fast response measured in milliseconds

• Ideal for transmission between nerves

• Binding of messenger leads directly to ion flows

across cell membrane

• Ion flow = secondary effect (signal transduction)

• Ion concentration within cell alters

• Leads to variation in cell chemistry

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G- Proteins ?

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 G-Proteins
• A family of membrane proteins that exist in an
inactive (GDP) and an active (GTP) state
• So-named because they bind GTP, displacing
GDP
• Work with many receptors
• Both Stimulate and inhibit hormone signals
• GTP is a time-bomb slowly ticking
• When GTP is hydrolyzed to GDP, stimulation is
stopped
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2. G-PROTEIN-COUPLED RECEPTORS

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 G-PROTEIN-COUPLED RECEPTORS…

Extracellular
loops NH2

N-Terminal chain

Transmembrane
Membrane VII VI V IV III II I helix

G-Protein
binding region
HO2C
Variable
C-Terminal chain intracellular loop Intracellular loops
Ligand binding site - varies depending on receptor type

Ligand
A B C D

A) Monoamines: pocket in TM helices

B) Peptide hormones: top of TM helices + extracellular loops

+ N-terminal chain

C) Hormones: extracellular loops + N-terminal chain

D) Glutamate: N-terminal chain

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 G-protein-coupled receptors…
• Receptor binds a messenger leading to an induced fit
• Opens a binding site for a signal protein (G-protein)
• G-protein binds, is destabilised then split

messenger

induced
fit

closed open

G-protein
split

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Activation of G Proteins

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 3. Tyrosine kinase - linked receptors
• Bifunctional receptor / enzyme

• Activated by hormones

• Overexpression can result in cancer

 Tyrosine kinase-linked receptors…

• Protein serves dual role - receptor plus enzyme

• Receptor binds messenger leading to an induced fit
• Protein changes shape and opens active site
• Reaction catalysed within cell
• Overexpression related to several cancers

messenger messenger

induced
fit

active site
closed closed
open

intracellular reaction
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 Tyrosine kinase-linked receptors…

Ligand binding region

Extracellular
NH2
N-terminal
chain

Hydrophilic
Cell membrane
transmembrane
region (a-helix)

Catalytic binding region

(closed in resting state)
Intracellular
C-terminal C O2 H
chain

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Reaction catalysed by tyrosine kinase

O Tyrosine O
N C kinase N C
Protein Mg++ Protein
Protein Protein

OH ATP ADP O P
Tyrosine
residue Phosphorylated
tyrosine
residue

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Growth Factors

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 Epidermal growth factor receptor (EGF- R)

EGF

Ligand binding
Cell and dimerisation Phosphorylation
membrane

HO OH PO OP
OH OH ATP OP OP
ADP
Inactive EGF-R Induced fit
monomers opens tyrosine kinase
active sites

Binding site for EGF

EGF - protein hormone - bivalent ligand
Active site of tyrosine kinase
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 Epidermal growth factor receptor (EGF- R)…

• Active site on one half of dimer catalyses phosphorylation

of Tyr residues on other half

• Dimerisation of receptor is crucial

• Phosphorylated regions act as binding sites for further

proteins and enzymes

• Results in activation of signalling proteins and enzymes

• Message carried into cell

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 Fibroblast Growth Factor

• What role does this growth factor play in embryonic

development?
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Transforming Growth Factor-β (TGF-β)
• How does TGF-β differ from FGF?

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 Insulin receptor (tetrameric complex)

Insulin

Phosphorylation
Cell
membrane
HO OH PO OP
ATP ADP OP OP
OH OH

Kinase active site

opened by induced fit

Insulin binding site

Kinase active site

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Growth hormone receptor
Tetrameric complex constructed in presence of
growth hormone
GH
GH binding
& Binding Activation and
dimerisation of kinases phosphorylation

GH receptors ATP ADP

(no kinase activity)
HO OH PO OP
kinases OH OH OP OP

HO Kinase active site

OH OH
OH opened by induced fit

Growth hormone binding site

Kinase active site
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 4. Intracellular receptors
• Chemical messengers must cross cell membrane
CO2H
• Chemical messengers must be hydrophobic

• Example-steroids and
steroid receptors
Steroid
binding region
Zinc

DNA binding region

(‘zinc fingers’)

H2N

Zinc fingers contain Cys residues (SH)

Allow S-Zn interactions

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 Intracellular receptor Mechanism

Co-activator
protein
Receptor

DNA
Messenger
Receptor-ligand Dimerisation
complex

Cell
membrane

1. Messenger crosses membrane 5. Complex binds to DNA

2. Binds to receptor 6. Transcription switched on or off
3. Receptor dimerisation 7. Protein synthesis activated or inhibited
4. Binds co-activator protein

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Thyroid hormone receptor
 Thyroid hormone receptor
• Thyroid hormone receptor (TR) heterodimerized
to the RXR
• In absence of a ligand, the TR is bound to a
corepressor protein
• Ligand binding to the TR causes a dissociation of
co-repressor and recruitment of co-activator
proteins, which in turn recruit additional
proteins (such as RNA polymerase) that are
responsible for the transcription of downstream
DNA into RNA, and eventually into protein that
results in a change in cell function.
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Other receptors

Bacteriorhodopsin & Rhodopsin Family

• Rhodopsin = visual receptor

•Implications for drug selectivity depending on
similarity (evolution)
•Bacteriorhodopsin structure used as ‘template’ for
other receptors
•Leads to model binding sites for drug design

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 Bacteriorhodopsin & Rhodopsin Family…

Common ance stor

Monoamines
muscarinic
alpha beta

Bradykinin, Opsins, Rhodopsins

Endothelins Angiotensin.Tachykinins
Interleukin-8 Rece ptor
ty pe s

Rece ptor
2 4 5 3 1 H1 H2 1 2A 2B 2C D4 D3 D2 D1A D1B D5 3 2 1 sub-ty pes

Muscarinic Histamine -Adren ergic Dopaminergic -Adren ergic

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Receptor types and subtypes of Rhodopsin Family
• Receptor types and subtypes not equally distributed
amongst tissues

• Target selectivity leads to tissue selectivity

Heart muscle b1 adrenergic receptors

Fat cells b3 adrenergic receptors
Bronchial muscle a1& b2 adrenergic receptors
GI-tract a1 a2 & b2 adrenergic receptors

End of Part A
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 Part A Recap
Describe how the following
receptors function
i. Ion channel receptors
ii. G-protein-coupled receptors
iii. Kinase-linked receptors
iv. Intracellular receptors

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 PART B

• Signal Transduction
• Second messengers

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