Cell Signaling

2. Components of
signaling pathways
1. The scheme of a typical pathway
2. Ligands
3. Receptors
4. Intracellular signal molecules
 General scheme of a typical signaling pathway 1.
Signal molecule (ligand; primary messenger)
Receptor protein

AC
Prim
ar y effe
Transducer ctor
Secondary effector
Second messenger
Other effectors Signaling proteins

Target proteins

Metabolic enzyme Cytoskeleton protein Changed shape

or movement
Altered metabolism Ion channel Transcription factor

Changed ion milieu Changed gene expression Cellular response

Cellular response
 2.
Signal molecules
1. Signals from the environment (light, sound)
2. Signals from other individuals (pheromones)
3. Signals from other cells
4. Signals from the same cell

NEURO- Hormones
TRANSMITTERS
Growth factors, neurotrophins
HORMONES
CYTOKINES
Components of extracellular matrix
Cytokines, chemokines
GROWTH
FACTORS
Neurotransmitters
 3.
Foreign signals
 4.

RECEPTORS
1. Intracellular receptors

2. Cell-surface receptors
a. Ion-channel -coupled receptors
b. G protein-coupled receptors

c. Enzyme-coupled receptors
The NO effects without receptor 5.

BLOOD VESSEL fibroblast cell

Smooth muscle cell
Endothelial cell

Acetylcholine lumen

Inhibits cGMP break-down

Smooth muscle
NO
NO synthase
Guanylate cyclase

NO NO NO
Arginine NO GTP cGMP
NO NO
NO - diffusion

Relaxed
Endothelial cell
Smooth muscle cell
Nitroglycerin → NO production
Viagra → inhibiton of cGMP degradation
 6.
Nobel Price for the discovery of the role of NO
(1998)

Robert F Furchgott Luise J Ignarro Ferid Murad

 7.
Intracellular receptor
Cell membrane

Hydrophobic signal molecule cytoplasm

receptor

transcription factor
Carrier protein

DNA
Nucleus
 8.
Intracellular receptors
(Nuclear receptors)
transcription

Nuclear receptor superfamily

Estrogen receptor
Cortisol receptor
Progesterone receptor
Vitamin D receptor
Thyroid hormone receptor
Retinoic acid receptor
 9.
Steroid hormone activation
inhibitor (hsp90)
steroid

glucocorticoid receptor

Nucleus

DNA GRE

cytoplasm

GRE:
GRE:glucocorticoid response
glucocorticoid response element
element
 10.
Activation by steroid hormones
1. Early primary response to steroid hormone

Steroid
hormone Receptor

2. Delayed secondary response to steroid hormone

Primary-response genes Secondary-response genes

DNA

Primary-response proteins
shut off primary-response turn on secondary-response
genes genes
Primary-response proteins
 11.
Cell-surface receptor
1. Ion channel-coupled receptors Cell membrane
2. G protein-coupled receptors
3. Enzyme-coupled receptors
Cytoplasm

receptor

Hydrophilic signal molecule

Nucleus
 12.
+
+ Receptors
+ + +
+ +
+ + + + Signal molecule
2. G protein-coupled receptor
Signal molecule
+

+
activated  subunit

CLOSED + OPEN

1. Ionotropic receptor + +
(ligand-activated ion channel) activated  complex
+ +
3a. Enzyme-coupled receptor 3b. Enzyme-coupled receptor
(the receptor itself is an enzyme) (the receptor binds to an enzyme)
Signal molecule
dimer signal molecule

activated receptor

Inactive Active
receptor tyrosine kinase receptor tyrosine kinase Inactive enzyme Activated enzyme
 13.
G protein-coupled receptors
Types: Gs, Gi, G0
s: stimulatory
i: inhibitory 1994
0: others
G protein
G protein-coupled
receptor
Alfred G. Gilman Martin Rodbell
Signal molecule
„For their discovery of G-proteins and the role of these proteins
in signal transduction in cells"

OLFACTION:
mammals: 1000 olfaction-related GPCR
human: only 391 active, ~600 pseudogene

activated  complex
activated  subunit
Adrenalin at least 9, acetylcholine 5, serotonin at least 14 different GPCR.
 14.
G-protein-linked receptor kinase
Activated Desensitized
receptor receptor

GRK Arrestin binds to

phosphorylates phosphorylated
activated receptor receptor
Arrestin

STOP! GRK
(G-protein-linked receptor kinase)
 15.
G protein
Inactive enzyme

activated  subunit

Activated enzyme

Cell response

Re-assembled G protein

inactivated  subunit
 16.
Receptor tyrosine kinase
subfamilies
 17.
Receptor tyrosine kinase &
activation of Ras
PDGF dimer
signal molecule

activated Ras
Inactive Ras

Signaling cascade
Inactive Activated Activated
receptor tyrosine receptor tyrosine kinase intracellular signaling molecules
kinase

Activation by cross-phosphorylation

PDGF : platelet-derived growth factor

 18.
Regulation of Ras activity
Inactive
Ras

GEF

GAP
Active
Ras

GEF: guanine nucleotide exchange factor

GAP: GTP-ase-activating protein (e.g. SOS1)
 19.
Intracellular signaling molecules

1. Small intracellular mediator

(second messengers)

2. Intracellular signaling proteins

 20.

Second messengers
1. cyclic nucleotides (cAMP, cGMP)
2. IP3, DAG
3. Ca2+
4. gases (NO, CO)
 21.
Signaling proteins
 Activation and inhibition of the 22.
components of signaling pathways

1. Phosphorylation
2. GDP-GTP exchange

3. Binding to a second messenger

4. Proteolytic cleavage
5. Dissociation of inhibitory protein (chaperon)
6. Ubiquitination
 SIGNAL IN
23.
Protein kinase
Inactive
enzyme
Phosphorylation
ON OFF Protein
phosphatase
Kinases
Active
enzyme
(1) Tyrosine kinases
(2) Serine/threonine kinases
SIGNAL OUT
Signal molecule

Activation through
Phosphorylation Receptor Activated relay molecule

Inactive
Inhibition through protein kinase
Active

Ph
1
Phosphorylation protein kinase

o
sp
1

ho
Inactive

ry
protein kinase

lati
Active
Activation through 2

on
protein kinase
Dephosphorylation 2

ca
sca
Inactive

d
protein kinase

e
Active
Inhibition through 3
protein kinase
Dephosphorylation 1
Inactive
protein
Active CELLULAR
protein RESPONSE
 24.
Phosphorylation cascade
Extracellular signals

Primary phosphorylation

Kinase 1 Kinase 2

Kinase 3
Secondary phophorylation

Kinase 4
 25.
Signaling by GTP-binding protein

SIGNALING BY GTP-BINDING PROTEIN

SIGNAL IN

P
GEF GD
Inactive

GA
P
GDP enzyme

GT
ON OFF
P

Active
enzyme

SIGNAL OUT
 26.
Inhibitors (e.g. chaperons)
For example: activation of cAMP dependent protein kinase - PKA

cAMP

Inactive catalytic subunit

Regulatory subunit Active catalytic subunit

(chaperon)
 27.
Proteolysis „chain reaction”
(caspase cascade in apoptosis)
 28.
Ca2+
Regulation of Ca level 2+
Na+ Na+/Ca2+
Ca2+ pump Ca2+ channel
exchanger (voltage-gated)

Cell membrane

cytoplasm

Ca2+ importer
Ca -binding molecule
2+
Ca pump
2+

Endoplasmic reticulum Mitochondrion

 29.
Activation of CaM-kinase-II
Ca2+-independent
Inactive

CaM-kinase-II

Ca2+
calmodulin Ca2+/calmodulin
Ca2+

Fully active Activated