Manifestation of Novel Social Challenges of the European Unionin the Teaching Material ofMedical Biotechnology Master’s Programmesat the University of Pécs and at the University of DebrecenIdentification number: TÁMOP-4.1.2-08/1/A-2009-0011
TYROSINE KINASE-LINKED RECEPTORS PART 2GROWTH FACTORS
Tímea Berki and Ferenc BoldizsárSignal transduction
Manifestation of Novel Social Challenges of the European Unionin the Teaching Material ofMedical Biotechnology Master’s Programmesat the University of Pécs and at the University of DebrecenIdentification number: TÁMOP-4.1.2-08/1/A-2009-0011
TÁMOP-4.1.2-08/1/A-2009-0011
Growth factors (GFs)• Small molecular weight soluble
mediators• They control:
1Proliferation2Survival3Metabolism4Tissue differentiation
• Important implication in tumors• Cytokines – growth factors
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Growth factors (GFs) – HistoryHow to propagate cells under in vitro conditions (turning of the 19th-20th century)?• Rous: experiments with chicken tumor (sarcoma) cells (RSV)• Carrel: In simple buffered salt solution the cells did not
proliferate – trials with diluted plasma/serum• Temin and Dulbecco: precise requirements for tissue culturing:
– Reduced serum need of tumor cells – enhanced capacity of tumor cells to respond to proliferation signals
– Serum supported cell growth rather than plasma – PDGFRita Levi-Montalcini, Stanley Cohen – NGF and EGF
Transplantation of an actively growing mouse tumor into chicken induced the great amplification of nerve fibres into the tumor mass
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Growth factorsFactor Principal source Primal activity Comments
PDGFPlatelets, endothelial cells, placenta
Promotes proliferation of connective tissue, glial and smooth muscle cells
Two differentprotein chains from 3 distinct dimer forms; AA, AB and BB
EGFSubmaxillary gland, Brunners gland
Promotes proliferation of mesenchymal, glial and epithelial cells
TGF-α Common in transformed cellsMay be important for normal wound healing
Related to EGF
FGFWide range of cells; protein is associated with the ECM
Promotes proliferation of many cells; inhibits some stem cells; induces mesoderm to form in early embrios
At least 18 family members, 5 distinct receptors
NGFMast cells, eosinophils, bone marrow stromal cells, keratinocytes
Promotes neurite outgrowth and neural cell survival
Member of a family of proteins termed neutrophins that promote proliferation and survival of neurons; neutrophin receptors are a class of related proteins first identified as proto-oncogenes? TrkA („trackA”), TrkB, TrkC
Erythropoietin Kidney Promotes proliferation of erythrocytes
TGF-β Activated Th1 cells (T-helper) and natural killer (NK) cells
Anti-inflammatory (suppresses cytokine production and class II MHC expression), promotes wound healing, inhibits macrophage and lymphocyte proliferation
At least 100 different family members
IGF-1 Primarily liver Promotes proliferation of many cell typesRelated to IGF-2 and proinsulin, also called somatomedin C
IGF-2 Variety of cellsPromotes proliferation of many cell types primarily of fetal origin
Related to IGF-1 and proinsulin
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Receptors with TK activity• Receptor Tyrosine Kinases (RTK)
e.g. PDGF, insulin, EGF, VEGF and FGF receptors
• Tyrosine-Kinase Associated ReceptorsReceptors that associate with proteins that have tyrosine kinase activity (Cytokine Receptors)
• Receptor Tyrosine Phosphatases e.g. CD45 protein of T cells and macrophages
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“Complete” and “incomplete” receptor tyrosine kinase
“Complete” receptor tyrosine kinase “Incomplete” receptor
Cytoplasmicnon-receptor
tyrosine kinase
Adaptor
Signal Signal
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Receptor tyrosine kinase (RTK) families• 90 unique Tyr kinases in the human genome, 58 are RTKs• Growth factor, cytokine and hormone receptors• Classes:
I EGFR family (ErbB) X LTK family
II Insulin rec. family XI TIE familyIII PDGF family XII ROR familyIV FGF family XIII DDR familyV VEGF family XIV RET familyVI HGF family (c-
Met) XV KLG familyVII Trk family XVI RYK familyVIII Eph family XVII MuSK familyIX AXL family
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Kinase-phosphatase balancePhosphorylase kinase (ser/thr
kinase)
PP1c (ser/thr phosphatase)
Phosphorylase b
Phosphorylase b
Phosphorylase a P
Phosphorylase a P
Inactive ActiveP
ATP ADP
CD45 (tyr phosphatase)
Csk (tyr kinase)ADP ATP
Inactive p56Lck
P Y505
Y394
Primed p56Lck
Y505
Y394
Active p56Lck
P Y394
P
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Fibronectin IIILeucine-rich
Cysteine-richAcid-box
KinaseIG-like
VEGFR1VEGFR2VEGFR3
PDGFRPDGFRβCSF1RKitKit2
Ryk TorsoEGFRErbB2ErbB3ErbB4
MetRonSea
TrkATrkBTrkC
INSRIGF1RIRR
AxlMerSky
EphEckEekErkElkEhk1Ehk2SekHekHek11Cek-9Myk-1Myk-2
RosFGFR1FGFR2FGFR3FGFR4
TieTie2
DDR Ret TorpedoRor1Ror2
LtkAlk
EGF-likeCadherin
Factor VIII-likeGlicyne-rich Kringle
C1r-like
Growth factor receptors
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Receptor-like PTPs (21)
CD45(RC)
R1/R6
PTP(RM)PTP(RK)PTP(RT)PTP(RU)
R2B
LAR(RF)PYP(RS)PTP(RD)
R2A
PTP(RA)PYP(RE)
R4
PTP(RG)PYP(RZ1)
R5
PTPβ(RB)DEP1(RJ)
SAP1(RH)
GLEPP1(RO)
PTPS31(RP)
R3
PCPTP1(RR)STEP(N5)
R7
IA2(RN)IA2β
(RN2)
R8
RGDS motif
Proline-richMAM domain
SEC14 domainFibronectin III
FERM domainIG-like
BRO-1 homologyGlycosylated
PDZ domainCadherine-like
Histidine domainCarbonic anhydrase-like
Kinase-interacting domainPTP domain
Src homology 2PTP pseudo-phosphatasedomain
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Nontransmembrane PTPs (17)
HDPTP(N23)
NT8
MEG2(N9)
NT3
HePTP(N7)
PTPH1(N3)
MEG1(N14)
NT5
SHP1(N6)SHP2(N11)
NT2
PTPBAS(N13)
NT7
PTPD1(N21)
PTPD2β(142)
NT6
PTP1B(N1)
TCPTPβ(N2)
NT1
BDP1(N18)
PTP-PEST(N12)LYP
(N220)
NT4
PTPTyP(N20)
NT9
RGDS motif
Proline-richMAM domain
SEC14 domainFibronectin III
FERM domainIG-like
BRO-1 homologyGlycosylated
PDZ domainCadherine-like
Histidine domainCarbonic anhydrase-like
Kinase-interacting domainPTP domain
Src homology 2PTP pseudo-phosphatasedomain
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Growth factor receptors and tyrosine phosphorylation
p120Ras-Gap
PLC
PhosphotyrosineY559Y581Y716Y741Y751Y771
Y1009Y1021
Phosphotyrosine Y992Y1045Y1068Y1086Y1148Y1173
PDGFR
PP
PPP
PP
PPP
PP
PP
EGFR
PPPPP
PPPP
P
Shc SLP76
Abl
CblPI3K
IRS-1
GRB2 SRCSHP-1
STAT1
Crk
Nck
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Receptor tyrosine kinase (RTK) signaling
Proliferation MigrationSurvival Cell cycle progression
Transcription
RTK
Ligand
P
PP
PP
PP
P
Dimerization
Src
SOSGRB2Ras
RafErk
PKC
PLC
STAT
JAK
Akt
PI3KPDK1
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GF receptor signaling pathways I
TargetsPIP2 PIP3
TargetsTargets
AktPDK1
PIP3
SOS Ras
Targets
Erk
Targets
GRB2GRB2
Shp2
Shp2
GRB2
GRB2
GRB2
PI3K
RTK
Ligand
P P
P P
P
P
P
PP
Plasma membrane
P
PP
PP
PP
P
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GF receptor signaling pathways II
RTK
Ligand
DAG
IP3PIP2
PKC
Ca2+
Cbl
PPLC PLC C2PH SH3SH2 SH2
Plasma membrane
CytoplasmP
PP
PP
PP
P
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Overview of EGF signaling
EGFR
JNK
SOS
Ras
GRB2
C-Fos
Raf
MAPKK
AP1
MAPK
PAK1
NckGRB2
Gab1
Shp2
WASP
Src
Shc
Bad FKHR
CREB
RSK2 Junp53
MAPK JNKp38
Cdc42/Rac
Vav2
EGF
CytoskeletonCell cycle
Apoptosis
STAT1 STAT3
Target genes
ADAM
HB-EGF
PTP
Rac
H2O2
NADPHsynthesis Gab1
PI3K
PIP3
E2Ub
Targets
DOK
Akt PDK1
Cbl
MKK2 MKK4
MEKK MEKK4
Rac
FAK
CAS
Paxillin
Src
Targets
Ca2+
PKC
DAG
PLC
IP3
MAPK
RasGAP
+
- -
- -
-
+
-
+
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General characteristics of GF signaling
Diverse input signals(Multiple RTKs)
Conserved coreprocesses
Diverse ouput events(transcriptional responses,cytokeletal changes, etc)
System control+
- +
+
-
Input layer
Output layer
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Different GF receptors use the same signaling pathways
PDGF-C
Cell survival Proliferation Apoptosis resistance
Metastasis Angiogenesis
P
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
PP
P
EGFR Her2 Her3 Her4 VEGFR1 VEGFR2 VEGFR3 PDGFR-a PDGFR-b c-kit
EGFTGF
β-cellulinAmphiregulin
HB-EGFNo specific
ligands Heregulins
β-cellulinNRG2NRG3 VEGF-B VEGF-A VEGF-C VEGF-D PDGF-A
PDGF-BPDGF-D SCF
SOSGRB2 Ras Rac RhoCDC42
MEK1/2
Erk
Raf
ERK pathway
MKK4/7
JNK
MEKK
JNK pathway
MKK3/6
p38
Tak
p38 pathway
Akt
mTor
PI3K
EverolimusImatinibTrastuzumabLeflunomideLapatinibGefitinibErlotinibPanitumumabSorafenib
CetuximabBevacizumabVandetanibSunitinibEnzastaurinPazopanibMotesanibMidostaurinTemsirolimusSirolimus
P
PP
PP
P
P
P
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Natriuretic peptide signaling
↑NP degradation↓cAMP?↑ IP3?
↑ Vasorelaxation↑ Diuresis, natriuresis↓ Renin, aldosterone↓ Cell proliferation↓ Cardiac fibrosis
↑ Vasorelaxation↓ Cell proliferation↑ Long bone gowth
Kinase homology domain
Plasma membrane
Ligand binding domain
Receptor
Hinge region
Guanylyl cyclase domain
Physiologic response
Natriuretic peptide
NPR-C NPR-A(GC-A)
NPR-B(GC-B)
ANP BNP CNP
cGMPGTP cGMPGTP
PPPPPP
PPPPP
PPPPP
PPPPPP
Natriureticpeptide
Hormone bound
Active
Desensitised
Kinase
Phosphatase
ATP
ATP
cGMPGTP
P P
P
P P
P P
Basal
ATP