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

+

- -

- -

-

+

-

+

TÁMOP-4.1.2-08/1/A-2009-0011

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

TÁMOP-4.1.2-08/1/A-2009-0011

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