Manifestation of Novel Social Challenges of the European Union

in the Teaching Material of

Medical Biotechnology Master’s Programmes

at the University of Pécs and at the University of Debrecen Identification number: TÁMOP-4.1.2-08/1/A-2009-0011

TYROSINE KINASE-

LINKED RECEPTORS

PART 2

GROWTH FACTORS

Tímea Berki and Ferenc Boldizsár

Signal transduction

Manifestation of Novel Social Challenges of the European Union

in the Teaching Material of

Medical Biotechnology Master’s Programmes

at the University of Pécs and at the University of Debrecen Identification 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:

1 Proliferation

2 Survival

3 Metabolism

4 Tissue differentiation

• Important implication in tumors

• Cytokines – growth factors

TÁMOP-4.1.2-08/1/A-2009-0011

Growth factors (GFs) – History

How 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 – PDGF

Rita 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

TÁMOP-4.1.2-08/1/A-2009-0011

Growth factors

Factor Principal source Primal activity Comments

PDGF Platelets, 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

EGF Submaxillary gland, Brunners gland

Promotes proliferation of mesenchymal, glial and epithelial cells

TGF-α Common in transformed cells May be important for normal wound healing

Related to EGF

FGF Wide 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

NGF Mast 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 types Related to IGF-2 and proinsulin, also called somatomedin C

IGF-2 Variety of cells Promotes proliferation of many cell types primarily of fetal origin

Related to IGF-1 and proinsulin

TÁMOP-4.1.2-08/1/A-2009-0011

Receptors with TK activity

• Receptor Tyrosine Kinases (RTK)

e.g. PDGF, insulin, EGF, VEGF and FGF receptors

• Tyrosine-Kinase Associated Receptors

Receptors that associate with proteins that have

tyrosine kinase activity (Cytokine Receptors)

• Receptor Tyrosine Phosphatases

e.g. CD45 protein of T cells and macrophages

TÁMOP-4.1.2-08/1/A-2009-0011

“Complete” and “incomplete” receptor

tyrosine kinase

“Complete” receptor tyrosine kinase “Incomplete” receptor

Cytoplasmic

non-receptor

tyrosine kinase

Adaptor

Signal Signal

TÁMOP-4.1.2-08/1/A-2009-0011

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 family

III PDGF family XII ROR family

IV FGF family XIII DDR family

V VEGF family XIV RET family

VI HGF family (c-Met) XV KLG family

VII Trk family XVI RYK family

VIII Eph family XVII MuSK family

IX AXL family

TÁMOP-4.1.2-08/1/A-2009-0011

Kinase-phosphatase balance

Phosphorylase kinase (ser/thr

kinase)

PP1c (ser/thr phosphatase)

Phosphorylase b

Phosphorylase b

Phosphorylase a P

Phosphorylase a P

Inactive Active

P

ATP ADP

CD45 (tyr phosphatase)

Csk (tyr kinase)

ADP ATP

Inactive p56Lck

P Y505

Y394

Primed p56Lck

Y505

Y394

Active p56Lck

P Y394

P

TÁMOP-4.1.2-08/1/A-2009-0011

Fibronectin III

Leucine-rich

Cysteine-rich

Acid-box

Kinase

IG-like

VEGFR1

VEGFR2

VEGFR3

PDGFR

PDGFRβ

CSF1R

Kit

Kit2

Ryk Torso EGFR

ErbB2

ErbB3

ErbB4

Met

Ron

Sea

TrkA

TrkB

TrkC

INSR

IGF1R

IRR

Axl

Mer

Sky

Eph

Eck

Eek

Erk

Elk

Ehk1

Ehk2

Sek

Hek

Hek11

Cek-9

Myk-1

Myk-2

Ros FGFR1

FGFR2

FGFR3

FGFR4

Tie

Tie2

DDR Ret Torpedo Ror1

Ror2

Ltk

Alk

EGF-like

Cadherin

Factor VIII-like

Glicyne-rich Kringle

C1r-like

Growth factor receptors

TÁMOP-4.1.2-08/1/A-2009-0011

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-rich MAM domain

SEC14 domain Fibronectin III

FERM domain IG-like

BRO-1 homology Glycosylated

PDZ domain Cadherine-like

Histidine domain Carbonic anhydrase-like

Kinase-interacting domain PTP domain

Src homology 2 PTP pseudo-phosphatase

domain

TÁMOP-4.1.2-08/1/A-2009-0011

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-rich MAM domain

SEC14 domain Fibronectin III

FERM domain IG-like

BRO-1 homology Glycosylated

PDZ domain Cadherine-like

Histidine domain Carbonic anhydrase-like

Kinase-interacting domain PTP domain

Src homology 2 PTP pseudo-phosphatase

domain

TÁMOP-4.1.2-08/1/A-2009-0011

Growth factor receptors and tyrosine

phosphorylation

p120 Ras-Gap

PLC

Phosphotyrosine Y559

Y581

Y716

Y741

Y751

Y771

Y1009

Y1021

Phosphotyrosine Y992

Y1045

Y1068

Y1086

Y1148

Y1173

PDGFR

P

P

P

P

P

P

P

P

P

P

P

P

P

P

EGFR

P

P

P

P

P

P

P

P

P

P

Shc SLP7

6

Abl

Cbl PI3K

IRS-1

GRB2 SRC SHP-

1

STAT1

Crk

Nck

TÁMOP-4.1.2-08/1/A-2009-0011

Receptor tyrosine kinase (RTK)

signaling

Proliferation Migration Survival Cell cycle progression

Transcription

RTK

Ligand

P

P

P

P

P

P

P

P

Dimerization

Src

SOS GRB2

Ras

Raf

Erk

PKC

PLC

STAT

JAK

Akt

PI3K

PDK1

TÁMOP-4.1.2-08/1/A-2009-0011

GF receptor signaling pathways I

Targets PIP2 PIP3

Targets Targets

Akt PDK1

PIP3

SOS Ras

Targets

Erk

Targets

GRB2

GRB2

Shp2

Shp2

GRB2

GRB2

GRB2

PI3K

RTK

Ligand

P P

P P

P

P

P

P

P

Plasma membrane

P

P

P

P

P

P

P

P

TÁMOP-4.1.2-08/1/A-2009-0011

GF receptor signaling pathways II

RTK

Ligand

DAG

IP3 PIP2

PKC

Ca2+

Cbl

P

PLC PLC C2 PH SH3 SH2 SH2

Plasma membrane

Cytoplasm P

P

P

P

P

P

P

P

TÁMOP-4.1.2-08/1/A-2009-0011

Overview of EGF signaling

EGFR

JNK

SOS

Ras

GRB2

C-Fos

Raf

MAPKK

AP1

MAPK

PAK1

Nck

GRB2

Gab1

Shp2

WASP

Src

Shc

Bad FKHR

CREB

RSK2 Jun p53

MAPK JNK p38

Cdc42 /Rac

Vav2

EGF

Cytoskeleton Cell cycle

Apoptosis

STAT1 STAT3

Target genes

ADAM

HB-EGF

PTP

Rac

H2O2

NADPH

synthesis 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

Ras GAP

+

- -

- -

-

+

-

+

TÁMOP-4.1.2-08/1/A-2009-0011

General characteristics of GF

signaling

Diverse input signals

(Multiple RTKs)

Conserved core

processes

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

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

P

EGFR Her2 Her3 Her4 VEGFR1 VEGFR2 VEGFR3 PDGFR-a PDGFR-b c-kit

EGF

TGF

β-cellulin

Amphiregulin

HB-EGF

No specific

ligands Heregulins

β-cellulin

NRG2

NRG3 VEGF-B VEGF-A VEGF-C VEGF-D PDGF-A PDGF-B

PDGF-D SCF

SOS GRB2 Ras Rac Rho CDC42

MEK1/2

Erk

Raf

ERK pathway

MKK4/7

JNK

MEKK

JNK pathway

MKK3/6

p38

Tak

p38 pathway

Akt

mTor

PI3K

Everolimus

Imatinib

Trastuzumab

Leflunomide

Lapatinib

Gefitinib

Erlotinib

Panitumumab

Sorafenib

Cetuximab

Bevacizumab

Vandetanib

Sunitinib

Enzastaurin

Pazopanib

Motesanib

Midostaurin

Temsirolimus

Sirolimus

P

P

P

P

P

P

P

P

TÁMOP-4.1.2-08/1/A-2009-0011

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

cGMP GTP cGMP GTP

P P

P P P P

P P P P P

P P P

P P

P P P P

P P

Natriuretic

peptide

Hormone bound

Active

Desensitised

Kinase

Phosphatase

ATP

ATP

cGMP GTP

P P

P

P P

P P

Basal

ATP