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 /1/A

ENZYME-LINKED RECEPTORS 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 /1/A

TÁMOP /1/A Receptors with enzyme activity Receptor Tyrosine Kinases (RTK) –e.g. PDGF, insulin, EGF, VEGF and FGF receptors Receptor Tyrosine Phosphatases –e.g. CD45 (cluster determinant-45) protein of T cells and macrophages Receptor Guanylate Cyclases –e.g. natriuretic peptide receptors Receptor Serine/Threonine Kinases –e.g. activin and TGF-β receptors Tyrosine-Kinase Associated Receptors: receptors that associate with proteins that have tyrosine kinase activity (Cytokine Receptors)

TÁMOP /1/A I Receptor classes I ClassExamplesStructural Features of Class IEGF receptor, NEU/HER2, HER3Cysteine-rich sequences IIInsulin receptor, IGF-1 receptor Cysteine-rich sequences; characterized by disulfide-linked heterotetramers IIIPDGF receptors, c-KitContain 5 immunoglobulin-like domains; contain the kinase insert IVFGF receptors Contain 3 immunoglobulin-like domains as well as the kinase insert; acidic domain V Vascular endothelial cell growth factor (VEGF) receptor Contain 7 immunoglobulin-like domains as well as the kinase Insert domain VI Hepatocyte growth factor (HGF) and scatter factor (SF) receptors Heterodimeric like the class II receptors except that one of the two protein subunits is completely extracellular. The HGF receptor is a proto-oncogene that was originally identified as the MET oncogene VII Neurotrophin receptor family (TRKA, TRKB, TRKC) and NGF receptor Contain no or few cysteine-rich domains; NGFR has leucine rich domain

TÁMOP /1/A II Receptor classes II LigandReceptorsLigand TrapsAccessory ReceptorsSmads TGF-  TbR-II TbR-I ALK5 Latency-associated polypeptide (LAP) Decorin a 2 -macroglobulin Betaglycan Smad2 Smad3 Smad4 Inhibin ActR-IIB ALK4 Betaglycan Smad2 Smad3 Smad4 Activin ActR-IIB ALK4 Follistatin Smad2 Smad3 Smad4 Nodal GDF1 Vg1 ActR-IIB ALK4 ALK7 DAN Cerberus Cripto Smad2 Smad3 Smad4 BMP2 BMP4 BMP7 BMPR-II ALK3 ALK6 DAN Cerberus Chordin/SOG Follistatin Noggin Smad1 Smad5 Smad8 Smad4 BMP4 GDF5 ActR-IIB ALK3 ALK6 Chordin/SOG Follistatin Noggin Smad1 Smad5 Smad8 Smad4 BMP7 ActR-IIB ALK2 Chordin/SOG Follistatin Noggin Smad1 Smad5 Smad8 Smad4 MIS AMHR-II ALK2 ALK3 ALK6 Smad1 Smad5 Smad8 Smad4 TGF-  TbR-II ALK1 Endoglin Smad1 Smad5 Smad8 Smad4

TÁMOP /1/A Introduction “Complete” receptor tyrosine kinase “Incomplete” receptor Cytoplasmic non-receptor tyrosine kinase Adaptor SignalSignal

TÁMOP /1/A Receptor tyrosine kinase (RTK) families 90 unique Tyr kinases in the human genome, 58 are RTKs Growth factor, cytokine and hormone receptors Classes:Classes: I EGFR family (ErbB) XLTK family II Insulin rec. family XITIE family IIIPDGF familyXIIROR family IVFGF familyXIIIDDR family VVEGF familyXIVRET family VI HGF family (c- Met) XVKLG family VIITrk familyXVIRYK family VIIIEph familyXVIIMuSK family IXAXL family

TÁMOP /1/A Fibronectin III Leucine-rich Cysteine-rich Acid-box Kinase IG-like VEGFR1 VEGFR2 VEGFR3 PDGFR  PDGFR β CSF1R Kit Kit2 RykTorsoEGFR 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 RosFGFR1 FGFR2 FGFR3 FGFR4 Tie Tie2 DDRRetTorpedoRor1 Ror2 Ltk Alk EGF-like Cadherin Factor VIII-like Glicyne-richKringle C1r-like Receptor tyrosine kinase family

TÁMOP /1/A Main steps of RTK activation 1 1Ligand binding 2 2Dimerization 3 3Autophosphorylation 4 4Signal complex (adapter proteins, kinases etc.)

TÁMOP /1/A Dimerization of GF receptors Plasma membrane Cytoplasm Juxtamembrane regionC-terminal regionActivation and catalytic loop (substrate precluding) Activation and catalytic loop (substrate accessible) P P P P P P P P PP ATPATP Dimerization

TÁMOP /1/A Autophosphorylation of RTKs p120 Ras-Gap p120 Ras-Gap PLC  Phosphotyrosine Y559 Y581 Y716 Y741 Y751 Y771 Y1009 Y1021Phosphotyrosine 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

TÁMOP /1/A Members of the initial signal complex Enzymes/transcription factors: Src/Syk family kinases, SHP-1, PLCg, Sos, Vav, RasGAP, STAT1 etc. Adaptors/regulators: Grb2, SLP-76, SOCS1, Nck, Shc, Crk-L, p85 etc. Adaptors/docking proteins: FRS2, IRS1, DOK1 etc.

TÁMOP /1/A Members of the signaling complex SRC PLC  PKC PI3K RAS RAF MEK MAPKs Plasma membrane SOS SHC GRB2 RSK FAK DifferentiationDifferentiation Transcriptional regulation Differentiation/GrowthDifferentiation/Growth Nucleus Y Y Y Y Y Y Growth factor/Hormone Receptor PTK Cytoplasm

TÁMOP /1/A Ras-MAPK pathway Elk-1 P P Plasma membrane Cytoplasm RAS SOS GRB2 Growth factor/Hormone Receptor PTK Acti ve RAS Acti ve RAS RAF MEK1 ERK2 Adaptor Guanine nucleotide exchange factor (GEF) GDP GTP P P MEK1 P P ERK2 P P P P P P SRE Elk-1 P P SRF Nucleus AP-1 P P cFos + ? Gene transcription

TÁMOP /1/A Parallel MAPK pathways ERK pathway p38 pathway JNK pathway Mitogens Stress/Cytokines Ligands and activators A-raf, B-raf, C-raf MEKK1, DLK, MLK2, ASK, TAK1, TAO, TAO2 MEKK1, DLK, MLK2, ASK, TAK1, TAO, TAO2 MAPKKK MEK1, MEK2 MEK3, MEK6 MEK4, MEK7 MAPKK ERK1, ERK2, ERK5 ERK3, ERK4 p38a, b, d JNK1, 2, 3 MAPK RSK1 RSK2 RSK3 RSK4 RSK1 RSK2 RSK3 RSK4 MSK1 MSK2 MSK1 MSK2 MNK1 MNK2 MK2 MK3 MK2 MK3 MK5B MK PDK1

TÁMOP /1/A MAPK-activated kinases (MKs) RSK (ribosomal S6 kinases) cytoplasmic (initiation factors of translation, apoptosis machinery, oestrogen rec., Sos), phosphorylated form can translocate to nucleus (ATF4, c-Fos, SRF) MSK (mitogen- and stress-activated kinases ) in nucleus (CREB, histone H3, HMGN1, ATF1) MNK (MAPK-interacting kinases) translation initiation complex

TÁMOP /1/A Turning-off the pathway Phosphatases PTP1B, SHP1/2, DEP1 Phosphorylation of GEF (Sos) decreases the affinity to the adapter (Grb2) Removal of cell surface receptors by endocytosis

TÁMOP /1/A MAPK/ERK in growth and differentation