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Tyrosine Phosphorylation: from Discovery to the Kinome and Beyond

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Tyrosine Phosphorylation: from Discovery to the Kinome and Beyond The 2007 Herbert Tabor - Journal of Biological Chemistry Lecture Tony Hunter Tyrosine phosphorylation: from discovery to the kinome and beyond ASBMB Annual Meeting April 28, 2007 So far 43 JBC papers and one submitted! The History of Protein Phosphorylation Protein kinase ATP ADP Protein P.Protein P Protein phosphatase The History of Protein Phosphorylation Phospho- Protein Phospho- Src serine in kinase tyrosine tyrosine proteins activity in fly eggs kinase 1932 1954 1964 1979 1900 1910 1920 1930 1940 1950 1960 1970 1980 1990 2000 Phospho Phospho- Phosph- cAMP Gleevec -protein tyrosine orylase depen- approved discovery synthesis kinase dent PK for CML 1906 1933 1959 1968 2001 The History of Protein Phosphorylation J. Biol. Chem. 2:127 (1906) Phospho- Protein Phospho- Src serine in kinase tyrosine tyrosine proteins activity in fly eggs kinase 1932 1954 1964 1979 1900 1910 1920 1930 1940 1950 1960 1970 1980 1990 2000 Phospho Phospho- Phosph- cAMP Gleevec -protein tyrosine orylase depen- approved discovery synthesis kinase dent PK for CML 1906 1933 1959 1968 2001 The History of Protein Phosphorylation J. Biol. Chem. 98:109 (1932) Phospho- Protein Phospho- Src serine in kinase tyrosine tyrosine proteins activity in fly eggs kinase 1932 1954 1964 1979 1900 1910 1920 1930 1940 1950 1960 1970 1980 1990 2000 Phospho Phospho- Phosph- cAMP Gleevec -protein tyrosine orylase depen- approved discovery synthesis kinase dent PK for CML 1906 1933 1959 1968 2001 The History of Protein Phosphorylation J. Biol. Chem. 100:583 (1933) Phospho- Protein Phospho- Src serine in kinase tyrosine tyrosine proteins activity in fly eggs kinase 1932 1954 1964 1979 1900 1910 1920 1930 1940 1950 1960 1970 1980 1990 2000 Phospho Phospho- Phosph- cAMP Gleevec -protein tyrosine orylase depen- approved discovery synthesis kinase dent PK for CML 1906 1933 1959 1968 2001 The History of Protein Phosphorylation J. Biol. Chem. 211:969 (1954) Phospho- Protein Phospho- Src serine in kinase tyrosine tyrosine proteins activity in fly eggs kinase 1932 1954 1964 1979 1900 1910 1920 1930 1940 1950 1960 1970 1980 1990 2000 Phospho Phospho- Phosph- cAMP Gleevec -protein tyrosine orylase depen- approved discovery synthesis kinase dent PK for CML 1906 1933 1959 1968 2001 The History of Protein Phosphorylation Burnett and Kennedy, J. Biol. Chem. 211:969 (1954) Phospho- Protein Phospho- Src serine in kinase tyrosine tyrosine proteins activity in fly eggs kinase 1932 1954 1964 1979 1900 1910 1920 1930 1940 1950 1960 1970 1980 1990 2000 Phospho Phospho- Phosph- cAMP Gleevec -protein tyrosine orylase depen- approved discovery synthesis kinase dent PK for CML 1906 1933 1959 1968 2001 The History of Protein Phosphorylation Phospho- Protein Phospho- Src serine in kinase tyrosine tyrosine proteins activity in fly eggs kinase 1932 1954 1964 1979 1900 1910 1920 1930 1940 1950 1960 1970 1980 1990 2000 Phospho Phospho- Phosph- cAMP Gleevec -protein tyrosine orylase depen- approved discovery synthesis kinase dent PK for CML 1906 1933 1959 1968 2001 Historic moments in the discovery of phosphotyrosine + Pi Pi 18/9/79 P.SER P.SER pH “1.9” “X” P.THR P.TYR P.THR _ | mT 14/6/79 | | | | acid LT mT mT IgH/Src in vivo acid protease v-Src increases P.Tyr levels in transformed chick cells uninfected v-Src-transformed + pH 1.9 + pH 3.5 32P-labeled control and RSV-transformed chick fibroblasts Hunter and Sefton, PNAS 77:1311 (1980) The History of Protein Phosphorylation Phospho- Protein Phospho- Src serine in kinase tyrosine tyrosine proteins activity in fly eggs kinase 1932 1954 1964 1979 1900 1910 1920 1930 1940 1950 1960 1970 1980 1990 2000 Phospho Phospho- Phosph- cAMP Gleevec -protein tyrosine orylase depen- approved discovery synthesis kinase dent PK for CML 1906 1933 1959 1968 2001 The History of Protein Phosphorylation Phospho- Protein Phospho- Src serine in kinase tyrosine tyrosine proteins activity in fly eggs kinase 1932 1954 1964 1979 1900 1910 1920 1930 1940 1950 1960 1970 1980 1990 2000 Phospho Phospho- Phosph- cAMP Gleevec -protein tyrosine orylase depen- approved discovery synthesis kinase dent PK for CML 1906 1933 1959 1968 2001 More than 29,000 papers on tyrosine kinases have been published since 1979! How many tyrosine kinases are there? 1. The finding that v-Src and c-Src phosphorylate tyrosine gave us the first tyrosine kinase in 1979 2. By the end of 1980 four tyrosine kinases were known (Src, Abl, EGF receptor, Fps/Fes) 3. By the end of 1990 over 50 tyrosine kinases had been identified in vertebrates and equal numbers of tyrosine kinases and serine kinases were known, leading to the prediction that there might be several 100 tyrosine kinases in a vertebrate genome and a total of over 1000 protein kinases 4. The complete human genome sequence reported in 2001 reveals that there are 90 tyrosine kinases (all the tyrosine kinases had been found by other means before the sequence was completed), out of a total of 518 protein kinases The History of Tyrosine Phosphorylation SH2 PI3 kinase PLCγ STATs domain associates as RTK are PTK identified with MT substrate substrates 1986 1987 1989 1992 v-Src MT + Src v-Erb is PTP1B PTP1B PTB Human protein + tyrosine derived is first and IRK domain kinome has kinase kinase from EGFR PTPase structures binds pY 90 PTKs 1977/8 1979 1984 1988 1994 1995 2002 1975 1980 1985 1990 1995 2000 2005 c-Src Abl + Bcr-Abl SH2 SH2 Inactive Gleevec gene EGFR fusion domain domain c-Src approved discovery are PTKs in CML binds P.Tyr structure structure for CML 1976 1980 1985 1990 1992 1997 2001 v-Src v-Src ∆C Cdc2 is Yersinia Grb2 kinase activation inhibited encodes SH2/SH3 sequence mechanism by P.Tyr PTPase adaptor 1980 1987 1989 1990 1992 Insulin receptor tyrosine kinase catalytic domain Hubbard et al. Nature 372:746 (1994) Receptor tyrosine kinases ? EGFR INSR PDGFRα FLT1 FGFR1 CCK4 MET TRKA AXL TIE EphA1 RYK DDR1 RET ROS LTK ROR1 MuSK RTK106 HER2 IGF1R PDGFRβ FLK1 FGFR2 RON TRKB MER TEK EphA2 DDR2 ALK ROR2 HER3 IRR CSF1R FLT4 FGFR3 TRKC SKY EphA3 HER4 KIT FGFR4 EphA4 FLK2 EphA5 EphA6 LET-23 DAF-2 F59F3.1 EGL-15 KIN15 T01G5.1 EphA7 C16B8.1 F11D5.3 C16D9.2 CAM-1 F59F3.5 KIN16 T17A3.8 EphA8 C25F6.4 F40G9.13 TKR-1 W04G5.6N EphB1 T10H9.2 C08H9.8 W04G5.6C EphB2 EphB3 F59F5.3 Y50D4B-4 Human RTKs EphB4 LMR1 M01B2.1 ZK938.5 EphB5 LMR2 58 (20 classes) R09D1.12 B0198.3 EphB6 LMR3 R09D1.13 F54F7.5 VAB-1 Worm RTKs 29 (11 classes) (B0252.1, F11E6.8, F40A3.5, R151.4, T148.1, T22B11.3 11 Unclassified Y38H6C.20, C24G6.2A, F08F1.1, F09A5.2, F09G2.1) NonreceptorNonreceptor protein-tyrosine tyrosine kinase kinasess (Myr) Y SH3 binding regio n DNA BD Actin BD Abl Y Fes/Fe r Y Syk/Zap70 JEF domai n Kinase-like domain Y Jak PH domain Y Tec Integrin-binding/JEF domain Y Focal adhesion-binding Fak Y Cdc42-binding Ack Myr Y Y Src Csk Y Srm PTK catalytic domain YY SH2 domain Rak/Fr k SH3 domain Y Y Brk/Si k What is tyrosine phosphorylation used for? 1. Growth factor signaling (and oncogenesis) 2. Cell adhesion, spreading, migration and shape 3. Cell differentiation in development 4. Cell cycle control 5. Gene regulation and transcription 6. Endocytosis and exocytosis 7. Insulin stimulation of glucose uptake 8. Angiogenesis (formation of new blood vessels) 9. Regulation of ion channels in nerve transmission TransmembraneTransmembrane signaling signaling by tyrosineby tyrosine phosphorylation phosphorylation EGF receptor Interferon receptor PDGF receptor T cell receptor NH 2NH 2 Ligand binding domain Out Plasma membrane NH 2 In Catalytic domain Transmembrane receptor Bimolecular receptor protein-tyrosine kinase protein-tyrosine kinase Receptor tyrosine kinase (RTK) signaling through SH2 and PTB P.Tyr-binding domain proteins Schlessinger, Cell 103:211 (2000) Stone age bioinformatics! Manual alignment - March 1985 (BB) Eukaryotic protein kinases have related catalytic domains Serine kinases PKA-C Cdc2 Tyrosine kinases c-Src EGFR Catalytic domain ~300 aa Protein kinase catalytic domain subdomains Hanks, Quinn and Hunter, Science 241:42 (1988) Structure of PKA catalytic subunit bound to PKI (5-24) and ATP N-lobe pT197 catalytic cleft C-lobe Knighton et al. Science 253:414 (1991) The birth of the kinome: a thousand and one protein kinases Src Phos K PKA Hunter, Cell 50:823 (1987) The first kinome Hunter and Plowman, TiBS 22:18 (1997) The first kinome tree Hunter and Plowman, TiBS 22:18 (1997) How many protein kinases are there? • S. cerevisiae (6217 genes) 130 (116) protein kinases (2.1%) but no bona fide tyrosine kinases • S. pombe (4624 genes) 128 (114) protein kinases (2.8%) but no TKs • C. elegans (19100 genes) 454 (434) protein kinases (2.4%) including 90 tyrosine kinases (20%) • D. melanogaster (13600 genes) 239 (223) protein kinases (1.8%) including 32 tyrosine kinases (14%) • H. sapiens (23,000 genes) 518 (478) protein kinases (2.2%) including 90 tyrosine kinases (16%) (chimpanzee kinome is essentially identical) • A. thaliana (26,800 genes) 1055 protein kinases (3.8%) but no tyrosine kinases (>630 RLK) M. brevicolli (unicellular choanoflagellate) has bona fide tyrosine kinases, SH2 domains and protein-tyrosine phosphatases (PTPs); the yeasts have PTPs, but no tyrosine kinases or SH2 domains ~2% of all genes in eukaryotes encode protein kinases Some more recent kinomes • Tetrahymena (27424 genes) 1069 PKs (3.8%) - no true TKs, but has TKLs and 83 two-component HisK. Has 630 PKs not assignable to known families or subfamilies, with 37 novel classes and 100s of unique PKs • Dictyostelium (12500 genes) 285 PKs (2.3%) 246 ePKs, including TKLs but no true TKs, plus 26 aPKs and 14 HisKs • Sea urchin (24000 genes) 353 PKs (1.5%) 329 ePKs, 24 aPKs (Strongylocentrotus) and 53 TKs, but no HisK.
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