Análisis Celular y Molecular de la Señalización Celular

TEMA 3
Receptores de la Superficie Celular
SURFACE RECEPTORS (TRANSMEMBRANE RECEPTORS)
 SURFACE RECEPTORS (TRANSMEMBRANE RECEPTORS)

Proteins that span the whole plasma membrane thickness with

one part of the receptor facing outside the membrane
(extracellular domains) and one part of the receptor facing
the interior of the cell (intracellular domains).
 SURFACE RECEPTORS (TRANSMEMBRANE RECEPTORS)

When the extracellular domain recognizes a specific ligand,

the whole receptor undergoes a conformation change that
affects the structure and properties of the intracellular
domain, allowing new actions.
 SURFACE RECEPTORS (TRANSMEMBRANE RECEPTORS)

Single receptors may transduce signals following ligand

binding, but most usually binding of the ligand provokes
interaction and/or association of several receptor molecules.
 COMPONENTS OF SURFACE RECEPTOR SIGNALLING PATHWAYS

1. Plasma membrane receptors

2. Cellular transducers of extracellular signals

3. Systems generating second messengers and changing metabolic environment

4. Intracellular second messengers

5. Calcium-binding proteins

6. Intracellular protein kinases

7. Transcription Factors
 COMPONENTS OF SURFACE RECEPTOR SIGNALLING PATHWAYS

• Plasma membrane receptors:

• 7-transmembrane-spanning domain receptors (7-TMS) or G-Protein-coupled receptors(GPCR)

• Catalytic receptors with 1 transmembrane-spanning domain:

• Receptor Tyrosine kinases

• Receptor Serine/Threonine kinases

• Receptors recruiting thyrosine kinases (Cytokine receptors)

• Receptor Guanyl cyclase

• Oligomeric ion channels

• Receptors for adhesion molecules (extracellular matrix or cell membrane)

• Cellular transducers of extracellular signals:

• Scaffolding proteins

• GTP-binding proteins (G-Proteins)

 COMPONENTS OF SURFACE RECEPTOR SIGNALLING PATHWAYS
• Systems generating second messengers and changing metabolic environment:
• Enzymes: Adenyl cyclase, Guanyl cyclase, Phospholipase C
• Ion channels
• Membrane transporters

• Intracellular second messengers:

• Cyclic nuceotides: cAMP, cGMP
• Inositol triphosphate (IP3)
• Diaciyl glycerol (DAG)
• Ca2+
• Nitric oxide

• Calcium-binding proteins:
• Calmodulin

• Intracellular protein kinases:

• Protein kinases of different types and roles

• Transcrìption Factors:

• Receptor-specific and/or Gene-specific

 PLASMA MEMBRANE RECEPTORS

• G-Protein-coupled receptors (GPCR) or Seven Transmembrane-

spanning domain receptors (7-TMS)
• Catalytic receptors:
• One Transmembrane-spanning domain (1-TMS):
• Receptor tyrosine kinases (RTKs)
• Receptor serine/threonine kinases
• Receptors recruiting tyrosine kinases (Cytokine receptors)
• Receptor Guanyl cyclase
• Oligomeric ion channels
• Receptors for adhesion molecules:
• Receptors for extracellular matrix ligands
• Receptors for cell membrane ligands
G-PROTEIN COUPLED RECEPTORS (GPCR)

2-adrenergic Molecule
receptors resembling
ligand

Cholesterol
 SEVEN TRANSMEMBRANE-SPANNING DOMAIN RECEPTORS (GPCR)

• Large family of plasma membrane receptors that

receive signals from many extracellular ligands:
• Photons
• Pheromones and odorant molecules
• Hormones
• Neurotransmitters
• They are coupled to G-Proteins which are able to
bind GTP and to act as the initial intracellular
transmitters of the received signal.
• Human genome codifies for about 900 GPCR of
which only about 200 have a known function.
• GPCR are involved in many diseases and are
therapeutic target of almost a half of modern
medical drugs.
SEVEN TRANSMEMBRANE-SPANNING DOMAIN RECEPTORS (GPCR)

2-adrenergic Molecule
receptors resembling
ligand

Plasma
membrane

Cholesterol
SEVEN TRANSMEMBRANE-SPANNING DOMAIN RECEPTORS (GPCR)
 SEVEN TRANSMEMBRANE-SPANNING DOMAIN RECEPTORS (GPCR)

• Receptors have 7 transmembrane helices

• Interact with heterotrimeric G-Proteins
• Gα binds GDP at rest
• Gβ and Gγ complete the heterotrimer

• Activation of receptor leads to:

• GDP release and GTP binding to Gα
• Dissociation of Gβ and Gγ

• Slow hydrolysis of GTP to GDP returns the

system to the resting state
SEVEN TRANSMEMBRANE-SPANNING DOMAIN RECEPTORS (GPCR)

G-proteins
GTPase Cycle
 SEVEN TRANSMEMBRANE-SPANNING DOMAIN RECEPTORS (GPCR)

• When a ligand binds to its GPCR:

• It provokes a conformational change in GPCR
that turns the receptor into a Guanine-nucleotide
exchanging factor (GEF).
• GPCR activates an associated G-protein,
allowing this protein to exchange the bound GDP
for a GTP molecule.
• The a-subunit of G protein bound to GTP
dissociates from subunits β and γ in order to
interact with other signalling or effector proteins
• GPCR initiate two different ways of signal
transduction:
• Cyclic AMP (cAMP) pathway
• Phosphatidil Inositol (PI) pathway
 SEVEN TRANSMEMBRANE-SPANNING DOMAIN RECEPTORS (GPCR)

G protein-coupled Plasma Activated Signaling Inactive

receptor membrane receptor molecule enzyme

GTP
GDP GDP
CYTOPLASM
G protein Enzyme GDP GTP
1 (inactive) 2

Activated
enzyme

GTP
GDP
Pi

3 Cellular response 4
RECEPTORS TYROSINE KINASES

2-adrenergic Molecule
receptors resembling
ligand

Cholesterol
RECEPTORS ASSOCIATED TO TYROSINE KINASES

The human
genome contains
518
protein kinase
genes (~1.7% of total)

Tyrosine
kinases account
for 17% of the
total
 RECEPTORS ASSOCIATED TO TYROSINE KINASES

Two major divisions of tyrosine kinases are

“receptors” and cytoplasmic “non-receptors”
 RECEPTORS ASSOCIATED TO TYROSINE KINASES

• Cell surface receptors obtain protein kinase activity in two

general ways:
• Receptor tyrosine kinases:
• Possess an intrinsic tyrosine kinase activity that is part of the
receptor protein.
• Examples include receptors for growth factors
• Non-receptor tyrosine kinases:
• Receptors lack self-contained kinase function and must recruit
intracellular protein kinases to the plasma membrane and
intracellular domains of receptors..
• Examples include receptors for cytokines
CATALYTIC RECEPTORS WITH 1 TMS DOMAIN: RECEPTOR TYROSINE KINASES

• Receptors with tyrosine kinase activity (RTK) are

high-affinity receptors for growth factors and protein
hormones.
• 58 out of 90 tyrosine kinases known are RTK.
• RTK include the insulin receptor and receptors for
several growth factors.
• RTK cross the membrane only once.
• Activation of RTK provokes phosphorylation of
tyrosine residues on its cytoplasmic domain and the
interaction with phospholipase Cg to induce the
synthesis of membrane-derived second
messengers.
CATALYTIC RECEPTORS WITH 1 TMS DOMAIN: RECEPTOR TYROSINE KINASES

RTK family classification and

structure/function
• Involved in diverse cellular responses:
– Cell division
– Differentiation
– Motility
• Subdivided into 10 subclasses based on
differences within extracellular, ligand-
binding domain of receptor
CATALYTIC RECEPTORS WITH 1 TMS DOMAIN: RECEPTOR TYROSINE KINASES

RTK family classification and

structure/function

• Four common structural features shared

among RTKs:
– Extracellular ligand-binding domain
– Single transmembrane domain
– Cytoplasmic tyrosine kinase domain(s)
– Regulatory domains
CATALYTIC RECEPTORS WITH 1 TMS DOMAIN: RECEPTOR TYROSINE KINASES
CATALYTIC RECEPTORS WITH 1 TMS DOMAIN: RECEPTOR TYROSINE KINASES

Receptor dimerization and auto-

transphosphorylation

• Ligand-induced RTK activation induces

receptor dimerization, leading to activation of
catalytic domains in each subunit
• Receptor auto-transphosphorylation:
– Further stimulates kinase activity
– Leads to phosphorylation of additional proteins
involved in receptor signalling pathway
– Provides “docking sites” for downstream signalling
proteins (Grb2, PI3-kinase, phospholipase Cg, etc.)
CATALYTIC RECEPTORS WITH 1 TMS DOMAIN: RECEPTOR TYROSINE KINASES

RTK dimerization and

autotransphosphorylation
 Figure 11.7c
CATALYTIC RECEPTORS WITH 1 TMS DOMAIN: TYROSINE KINASES
Signaling Ligand-binding site
molecule (ligand)
a helix in the Signaling
membrane molecule

Tyr Tyr Tyr Tyr Tyr

Tyrosines Tyr
Tyr Tyr Tyr Tyr Tyr
Tyr
Tyr Tyr Tyr Tyr Tyr
Tyr

CYTOPLASM Receptor tyrosine Dimer

kinase proteins
1 (inactive monomers) 2

Activated relay
proteins

Cellular
P Tyr P Tyr Tyr P
Tyr Tyr Tyr P response 1
Tyr Tyr P Tyr Tyr P P Tyr Tyr P
Tyr Tyr P Tyr Tyr P P Tyr Tyr P Cellular
6 ATP 6 ADP
response 2
Activated tyrosine Fully activated
kinase regions receptor tyrosine
(unphosphorylated kinase Inactive
dimer) (phosphorylated relay proteins
3 4
dimer)
RECEPTOR SERINE/THREONINE KINASES

2-adrenergic Molecule
receptors resembling
ligand

Cholesterol
 RECEPTOR SERINE/THREONINE KINASES

• Includes the receptor for Transforming Growth Factor

β (TGFβ) and other growth- and differentiation factors.
• Signaling begins with the binding of a ligand of the
2-adrenergic Molecule
TGFβ ligand superfamily to a TGF- type
receptors II receptor.
resembling
ligand

Cholesterol
 RECEPTOR SERINE/THREONINE KINASES

• Type II receptor is a serine/threonine receptor kinase, which

catalyzes the phosphorylation of the TGF- Type I receptor.
• In mammals there are 7 Type I and 5 Type II receptors.
2-adrenergic Molecule
• Each class of ligand binds to a specific
receptors type IIresembling
receptor.
ligand

Cholesterol
 RECEPTOR SERINE/THREONINE KINASES

• Type I receptor then

phosphorylates R-Smads,
which transduce the signal
2-adrenergic Molecule
into the nucleus receptors
via a resembling
ligand
complex of Smad and
Smad4.

• R-Smad/Smad4 complex
binds to various transcription
Cholesterol
factors that are regulated
developmentally.
RECEPTORS ASSOCIATED TO TYROSINE KINASES: CYTOKINE RECEPTORS
 RECEPTORS ASSOCIATED TO TYROSINE KINASES: CYTOKINE RECEPTORS

• Ligand binding to receptor activates the bound Janus Kinase

(JAK)
• JAK + ATP -> JAK-P + ADP
• Activated receptors are coupled to JAK-STAT pathways
• JAK-P phosphorylates STAT proteins (Signal Transducers and
Activators of Transcription)

• Phosphorylated STATs (P-STAT) form dimers that become

transcription factors:
• Transported to nucleus
• Bind to DNA
• Activate transcription
CATALYTIC RECEPTORS ASSOCIATED TO TYROSINE KINASES
CATALYTIC RECEPTORS ASSOCIATED TO TYROSINE KINASES
CATALYTIC RECEPTORS WITH 1 TMS DOMAIN: GUANYL CYCLASE
 CATALYTIC RECEPTORS WITH 1 TMS DOMAIN: GUANYL CYCLASE

• After ligand binding, receptors with guanyl cyclase activity

produce second messenger cGMP from GTP.

• cGMP is an activator of protein kinases, such as protein

kinase G (PKG).

• An example of such receptors is the receptor for the hormone

atrial natriuretic peptide.

GTP GMPc
CATALYTIC RECEPTORS WITH 1 TMS DOMAIN: GUANYL CYCLASE

Other intracellular Guanyl cyclases include

the intracellular receptor for Nitric Oxide (NO)
 TYPES OF SURFACE RECEPTORS: ION-CHANNEL RECEPTORS

1 2 3

Gate
closed Ions Gate Gate closed
Signaling open
molecule
(ligand)

Plasma
Ligand-gated membrane
ion channel receptor
Cellular
response
 TYPES OF SURFACE RECEPTORS: ION-CHANNEL RECEPTORS

• A ligand-gated ion channel receptor acts as a gate

when the receptor changes shape
• When a signal molecule binds as a ligand to the
receptor, the gate allows specific ions, such as Na+
or Ca2+, through a channel in the receptor
1 2 3

Gate
closed Ions Gate Gate closed
Signaling open
molecule
(ligand)

Plasma
Ligand-gated membrane
ion channel receptor
Cellular
response
 TYPES OF SURFACE RECEPTORS: ION-CHANNEL RECEPTORS

Neuronal Action Potentials

• At rest K+ channel is open;
– K+in > K+out Δψ = -60 mV
• Acetylcholine receptor channels open upon
binding ligand
• Na+ and Ca2+ can pass to cell interior
membrane depolarizes
• Prolonged ligand binding result in ligand-bound
closed state
• Acetyl choline is hydrolyzed by
acetylcholinesterase in synapse
• Voltage gated Na+ channels open (Na+ out =>
in) upon depolarization (amplify the signal)
• Voltage gated K+ channels open (K+ in => out)
membrane repolarizes locally
• Wave of depolarization and repolarization
(action potential) transmitted along the axon to
the next synapse
• Voltage gated Ca2+ channels allow Ca2+ influx
• Ca2+ triggers SNAP/SNARE membrane fusion
event that releases acetylcholine from
exocytotic vesicles into the synapse
 TYPES OF TRANSPORT PROTEINS AT PLASMA MEMBRANE

Gradients are
indicated by
triangles whose
apex points
towards smaller
concentration,
smalle r electric
potential or both.

Bombas
impulsadas por ATP Canales iónicos Transportadores

• Pumps: They use the energy released by ATP hydrolysis to transport small
molecules or specific ions against electrochemical gradients.
• Channels: They catalyze the movement of specific ions in favor of
electrochemical gradients.
• Transporters: They facilitate the movement of ions or small molecules
TYPES OF SURFACE RECEPTORS: ION-CHANNEL RECEPTORS

K+ CHANNEL
TYPES OF SURFACE RECEPTORS: ION-CHANNEL RECEPTORS
TYPES OF SURFACE RECEPTORS: ADHESION MOLECULES
TYPES OF SURFACE RECEPTORS:LIGANDS OF ADHESION MOLECULES
TYPES OF SURFACE RECEPTORS:LIGANDS OF ADHESION MOLECULES
TYPES OF SURFACE RECEPTORS: ADHESION MOLECULES
TYPES OF SURFACE RECEPTORS: INTEGRINS
TYPES OF SURFACE RECEPTORS: INTEGRINS
 TYPES OF SURFACE RECEPTORS: SYNDECANS

• Syndecans are single transmembrane domain

proteins that act as co-receptors, especially for G
protein-coupled receptors.
• They carry 3-5 heparan sulfate and chondroitin
sulfate chains, which allow for interaction with a
large variety of ligands:
• Fibroblast growth factors
• Vascular endothelial growth factor
• Transforming growth factor-beta
• Fibronectin
• Antithrombin-1
 TYPES OF SURFACE RECEPTORS: SYNDECANS

• Nearly all extracellular matrix molecules contain

binding sites for both integrins and syndecans.
• Full cell-adhesion response requires engagement of
both receptor types.
• Ligation of both integrin and syndecan receptors is
required for focal adhesion formation and migration.
• Perturbation of both receptor types leads to substantial
wound healing defects.
TYPES OF SURFACE RECEPTORS: SYNDECANS