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Protein Domains That Interact with Receptor Tyrosine Kinases: Structural Aspects

Protein Domains That Interact with Receptor Tyrosine Kinases: Structural Aspects. Ming-Ming Zhou, Ph.D. MSSM Department of Pharmacology Course on “Cell Signaling Systems” January 27, 2005. The Role of Protein Domains in Cell Signaling. Growth Factors. RTK. Ras. Ras. P. P. P. P. P.

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Protein Domains That Interact with Receptor Tyrosine Kinases: Structural Aspects

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  1. Protein Domains That Interact with Receptor Tyrosine Kinases: Structural Aspects Ming-Ming Zhou, Ph.D. MSSM Department of Pharmacology Course on “Cell Signaling Systems” January 27, 2005

  2. The Role of Protein Domains in Cell Signaling Growth Factors RTK Ras Ras P P P P P P GDP Shc GTP PTB CH P Sos Raf PLC- SH2 SH2 MEK PI(3)K SH3 SH3 Grb2 MAPK c-fos, c-myc, c-jun

  3. Protein Domains in Cell Signal Transduction PH SH2 SH3 WW PTB -pY-X-X-hy- -hy-X-N-P-X-(p)Y- Phospholipids -P-X-X-P-X- -P-P-X-Y- 14-3-3 EH PDZ . . . -N-P-F- -R-S-X-pS-X-P- -E-S/T-D/V-COOH-

  4. Ligand Recognition of the Shc PTB and SH2 Domains Protein pTyr pTyr Sites Peptides Shc SH2 Shc PTB Kd (M) Kd (M) Human TrkA, pY490 HIIENPQpYFSDA 280 0.053 Mouse Polyoma MT, pY250 SLLSNPTpYSVMRS 320 0.140 Human ErbB3, pY1309 SAFDNPDpYWHSRLF 230 0.020 Human ErbB2, pY1248 AENPEpYLGLDV 1290 N.D. Human EGFR, pY1086 GSVQNPVpYHNQPLN 220 5.30 Human EGFR, pY1148 SLDNPDpYQQDFF 1000 0.200 Human EGFR, pY1114 TAVGNPEpYLNTVQ 290 0.140 Human EGFR, pY992 VADADEpYLIPQQ 130 680 Human EGFR, pY1173 TAENAEpYLRVA 60 770 Human TCR  Chain, pY141 GHDGLpYQGLSTATK 50 >10000 Ref: Zhou et al (1995) JBC270, 31119-31123.

  5. Phosphotyrosine Binding Domains  SH2 Domain: ---pYxx--- PTB Domain: ---xNPxpY--- Q:  Ligand Specificity  Structural Versatility  Functional Diversity

  6. 3D Structue of the Shc SH2 Domain/TCR Peptide Complex G R A2 A BG7 pY D B L D6 BG A BG H D4  R B5 N A T E4 D R B5 EF V C3  C pY+3  F F I BG4 pY+3 L D5 pY E B C C  D’ C G BC  N TCR  chain peptide: GHDGLpYQGLSTATK Ref: Zhou et al (1995) PNAS92, 7784-7788.

  7. Structures of SH2 Domains Shc Lck D G A BG B BG pY A pY+3 pY pY+3 E F B D’ F C E C D’ N DE DE PLC-1 Syp BG BG pY pY pY+3 pY+3 F E F D’ E D’ DE DE

  8. Structures of SH2 Domains PLC-1 Syp Lck Shc

  9. 3D NMR Structure of the Shc PTB Domain/TrkA Complex Ref: Zhou et al (1995) Nature 378, 584-592.

  10. FXNPXpY Recognition by the Shc PTB Domain

  11. PTB Domain versus SH2 Domain Shc PTB Domain Shc SH2 Domain

  12. Biological Ligands of PTB Domains PTB Domain Binding Partners Binding Sites ShchTrkA, pY490HIIENPQpYFSDA mPolyoma MT, pY250 SLLSNPTpYSVMRS hErbB3, pY1309SAFDNPDpYWHSRL hEGFR, pY1148ISLDNPDpYQQDFF IRS-1hIR, pY960LYASSNPEpYLSAS hIL-4R, pY497LVIAGNPApYRSFS hIGFIR, pY950LYASVNPEpYFSAS NumbLNXGLDNPAYTSSV NAKGFSNMSFEDFP Peptide LibraryYIGPYL X11, Fe65, Ced-6bAPPGYENPTYKFFE

  13. 3D Structure of the IRS-1 PTB Domain/IL-4R Complex Ref: Zhou et al (1996) Nature Structural Biology3, 388-393.

  14. Ligand Recognition of the IRS-1 PTB domain

  15. Distinct Mechanisms for NPXpY Recognition IRS-1 PTB Domain Shc PTB Domain

  16. PTB & PH Domains: A Conserved Structural Fold A B C D

  17. SNT/FRS2 Binding Sites in Neurotrophic Receptors • FGFRs(juxtamembrane region): • KSDFH SQMAV HKLAK SIPLR RQVTS • NGFRs: • TrkA,pY960HIIENPQpYFSDA • TrkBPVIENPQpYFGIT • TrkCPVIENPQpYFRQG • RET, pY1062 • TWIENKLpYGRIS

  18. 3D Structure of the SNT-1 PTB Domain/FGFR1 Complex C C 8 a1 b’ b’ 7 8 5 4 6 5 3 C 6 N FGFR FGFR 7 C 1 N 2 2 a1 N 3 1 4 N Ref: Dhalluin et al (2000) Molecular Cell6, 921-929.

  19. SNT-1 versus IRS-1 PTB domains C R63 R78 a1 8 4 b’ 5 3 6 FGFR 7 2 N N 1 R227 C C pY IL-4R a1 R212 4 pY-3 5 3 N 6 7 C 2 pY-6 1 N pY-8 SNT-1 A B IRS-1

  20. Sequence Alignment of the SNT & IRS PTB Domains

  21. SNT-1 Truncation Effect on FGFR & TRK Binding 1 FGFR TrkB N C 7 2 N 6 5 b’ 3 Vector hFGFR1 TRKB 1 N 7 8 a1 a1 6 Leu-, Trp- His-, Leu-, Trp- Leu-, Trp- His-, Leu-, Trp- Leu-, Trp- His-, Leu-, Trp- 4 5 pY-5 2 C R63 R78 3 Vector 4 R63 R78 C SNT-1 PTB Domain pY C SNT-1 PTB Domain Db8 w/FGFR Yeast Two-Hybrid Binding Assay w/TrkB

  22. Unifying Features of PTB Domains Ref: Yan & Zhou (2002) FEBS Lett.513, 67-70.

  23. Structure & Function of the SH3 Domain A C B Ref: Zarrinpar (2003) Sci. STKE2003(179), re8

  24. RTKs: Activation by Receptor Dimerization Ref: Hubbard, et al (1998) JBC273, 11987-11990.

  25. Autoinhibition of Insulin and FGF Receptors IRK FGFR1K Unphosphorylated FGFR1K Tris-phosphorylated IRK (IRK3P) Unphosphorylated IRK (IRK0P) Tris-phosphorylated IRK (IRK3P) - Cis-inhibition & Trans-activation

  26. AutoRegulation & Activation of Src Tyrosine Kinase

  27. 3D Structure of Protein Tyrosine Kinase Hck Ref: Xu, et al (1997) Nature 385, 595-602; Sicheri, et al (1997) Nature385, 602-609.

  28. Rational Design of Nonpeptide Inhibitors of Src SH2 Domain Ref: Sundaramoorthi et al (2003) Biopolymers71, 717-729.

  29. 3D Structure of the Abl Kinase Domain/Drug Complex Ref: Daub, H. et al (2004) Nature Review Drug Discovery3, 1001-1010.

  30. Summary Structure-Function of Protein Domains in RTKs  Conserved 3D structural fold with divergent sequences  Conserved structural fold with different functions  Single protein domain with multiple functions  Cooperative regulation of RTK activity in cell signaling

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