RECEPTOR PHYSIOLOGY
Rei Kristoffer C. Ubiña, MD
Department of Physiology
THE ROLE OF THE RECEPTOR
Globular proteins
Located mostly in the cell membrane
Receivemessages 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
THE ROLE OF THE RECEPTOR
Nerve Nerve
Signal
Messenger
Receptor
Response
Nucleus
Cell Cell
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
THE ROLE OF THE RECEPTOR
• Receptors contain a binding site (hollow or 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
THE BINDING SITE
Messenger Induced fit Messenger
Messenger
Cell
Membrane Receptor Receptor Receptor
Cell Cell Cell
message
Message
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
THE BINDING SITE
Messenge
r 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
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
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
MAIN TYPES OF RECEPTORS
ION CHANNEL RECEPTORS
G-PROTEIN-COUPLED RECEPTORS
KINASE-LINKED RECEPTORS
INTRACELLULAR RECEPTORS
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 reactions within
cell
ION CHANNEL RECEPTORS
Hydrophilic
tunnel
Cell
membrane
MESSENGER
RECEPTOR
BINDING
ION
SITE
CHANNEL
(open)
MESSENGER
Induced fit
and opening
Lock of ion channel
Gate Cell Ion Ion Cell
Cell Ion Ion 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
GATING
Binding site
Receptor Messenger
Induced
Cell Cell
membrane fit membrane
‘Gating’
(ion channel
opens)
Five glycoprotein subunits
traversing cell membrane
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
GATING
Fast response measured in msec
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
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
G-PROTEIN-COUPLED RECEPTORS
• G-protein subunit activates membrane bound enzyme
• Binds to allosteric binding site
• Induced fit results in opening of active site
• Intracellular reaction catalysed
Enzyme Enzyme
active site active site
(closed) (open)
Intracellular
reaction
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
Bacteriorhodopsin & Rhodopsin Family
• Rhodopsin = visual receptor
• Many common receptors belong to this same family
• Implications for drug selectivity depending on similarity (evolution)
• Membrane bound receptors difficult to crystallise
• X-Ray structure of bacteriorhodopsin solved - bacterial protein similar
to rhodopsin
• Bacteriorhodopsin structure used as ‘template’ for other receptors
• Construct model receptors based on template and amino acid
sequence
• Leads to model binding sites for drug design
• Crystal structures for rhodopsin and b2-adrenergic receptors now
solved - better templates
Bacteriorhodopsin & Rhodopsin Family
Common ancestor
Monoamines
muscarinic
alpha beta
Bradykinin, Opsins, Rhodopsins
Endothelins Angiotensin.Tachykinins
Interleukin-8 Receptor
types
Receptor
2 4 5 3 1 H1 H2 1 2A 2B 2C D4 D3 D2 D1A D1B D5 3 2 1 sub-types
Muscarinic Histamine -Adrenergic Dopaminergic -Adrenergic
RECEPTOR TYPES AND SUBTYPES
• 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
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
Tyrosine kinase-linked receptors
Ligand binding region
Extracellular
N H2
N-terminal
chain
Hydrophilic
Cell membrane
transmembrane
region (a-helix)
Catalytic binding region
(closed in resting state)
Intracellular
C-terminal C O2H
chain
Reaction catalysed by tyrosine kinase
O Tyrosine O
N C kinase N C
Protein Protein Mg++ Protein Protein
OH ATP ADP O P
Tyrosine Phosphorylated
residue tyrosine
residue
Epidermal growth factor receptor (EGF- R)
EGF
Cell Ligand binding
and dimerisation Phosphorylation
membrane
HO OH PO OP
OH OH OP OP
ATP 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
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
Insulin receptor (tetrameric complex)
Insulin
Phosphorylation
Cell
membrane
HO OH PO OP
OH ATP ADP OP OP
OH
Kinase active site
opened by induced fit
Insulin binding site
Kinase active site
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
Intracellular receptors
• Chemical messengers must cross cell CO2H
membrane
• Chemical messengers must be hydrophobic
• Example-steroids and Steroid
steroid receptors binding region
Zinc
DNA binding region
(‘zinc fingers’)
H2 N
Zinc fingers contain Cys residues (SH)
Allow S-Zn interactions
Intracellular receptor mechanism
Co-activator
protein
Receptor
DNA
Messenger
Receptor-ligand Dimerisation
complex
Cell
membrane
. Messenger crosses membrane5. 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
