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PMID: 12773374 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Structural basis for SH3 domain-mediated high-affinity binding between Mona/Gads and SLP-76.

The EMBO journal ·Vol. 22 ·No. 11 ·2003-06-02 ·Pages 2571-82

Harkiolaki M, Lewitzky M, Gilbert RJ, Jones EY, Bourette RP, Mouchiroud G, Sondermann H, Moarefi I, Feller SM

Abstract

SH3 domains are protein recognition modules within many adaptors and enzymes. With more than 500 SH3 domains in the human genome, binding selectivity is a key issue in understanding the molecular basis of SH3 domain interactions. The Grb2-like adaptor protein Mona/Gads associates stably with the T-cell receptor signal transducer SLP-76. The crystal structure of a complex between the C-terminal SH3 domain (SH3C) of Mona/Gads and a SLP-76 peptide has now been solved to 1.7 A. The peptide lacks the canonical SH3 domain binding motif P-x-x-P and does not form a frequently observed poly-proline type II helix. Instead, it adopts a clamp-like shape around the circumfence of the SH3C beta-barrel. The central R-x-x-K motif of the peptide forms a 3(10) helix and inserts into a negatively charged double pocket on the SH3C while several other residues complement binding through hydrophobic interactions, creating a short linear SH3C binding epitope of uniquely high affinity. Interestingly, the SH3C displays ion-dependent dimerization in the crystal and in solution, suggesting a novel mechanism for the regulation of SH3 domain functions.

MeSH Terms
Adaptor Proteins, Signal Transducing Amino Acid Motifs Amino Acid Sequence Animals Binding Sites Carrier Proteins/chemistry,genetics,metabolism Crystallography, X-Ray Dimerization Humans Hydrogen Bonding In Vitro Techniques Mice Models, Molecular Molecular Sequence Data Phosphoproteins/chemistry,genetics,metabolism Phylogeny Protein Binding Recombinant Proteins/chemistry,genetics,metabolism Sequence Homology, Amino Acid Static Electricity src Homology Domains
Chemicals
Adaptor Proteins, Signal Transducing Carrier Proteins GRAP2 protein, human Mona protein, mouse Phosphoproteins Recombinant Proteins SLP-76 signal Transducing adaptor proteins
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Harkiolaki Maria
Cancer Research UK Cell Signalling Group and Weatherall Institute of Molecular Medicine, Oxford, UK.
Lewitzky Marc
Gilbert Robert J C
Jones E Yvonne
Bourette Roland P
Mouchiroud Guy
Sondermann Holger
Moarefi Ismail
Feller Stephan M
References (74)
74 references, click to expand
  1. The hematopoietic-specific adaptor protein gads functions in T-cell signaling via interactions with the SLP-76 and LAT adaptors.
    Curr Biol. 1999 Jan 28;9(2):67-75 PMID: 10021361
  2. Automated MAD and MIR structure solution.
    Acta Crystallogr D Biol Crystallogr. 1999 Apr;55(Pt 4):849-61 PMID: 10089316
  3. Further additions to MolScript version 1.4, including reading and contouring of electron-density maps.
    Acta Crystallogr D Biol Crystallogr. 1999 Apr;55(Pt 4):938-40 PMID: 10089341
  4. GrpL, a Grb2-related adaptor protein, interacts with SLP-76 to regulate nuclear factor of activated T cell activation.
    J Exp Med. 1999 Apr 19;189(8):1243-53 PMID: 10209041
  5. Grf40, A novel Grb2 family member, is involved in T cell signaling through interaction with SLP-76 and LAT.
    J Exp Med. 1999 May 3;189(9):1383-90 PMID: 10224278
  6. ESPript: analysis of multiple sequence alignments in PostScript.
    Bioinformatics. 1999 Apr;15(4):305-8 PMID: 10320398
  7. Automated protein model building combined with iterative structure refinement.
    Nat Struct Biol. 1999 May;6(5):458-63 PMID: 10331874
  8. A novel peptide-SH3 interaction.
    EMBO J. 1999 Oct 1;18(19):5300-9 PMID: 10508163
  9. The SH3 domains of endophilin and amphiphysin bind to the proline-rich region of synaptojanin 1 at distinct sites that display an unconventional binding specificity.
    J Biol Chem. 1999 Nov 5;274(45):32001-7 PMID: 10542231
  10. Lad, an adapter protein interacting with the SH2 domain of p56lck, is required for T cell activation.
    J Immunol. 1999 Nov 15;163(10):5242-9 PMID: 10553045
  11. The importance of being proline: the interaction of proline-rich motifs in signaling proteins with their cognate domains.
    FASEB J. 2000 Feb;14(2):231-41 PMID: 10657980
  12. Structure and dimerization of a soluble form of B7-1.
    Immunity. 2000 Jan;12(1):51-60 PMID: 10661405
  13. GCIP, a novel human grap2 and cyclin D interacting protein, regulates E2F-mediated transcriptional activity.
    J Biol Chem. 2000 Jul 7;275(27):20942-8 PMID: 10801854
  14. GRID: a novel Grb-2-related adapter protein that interacts with the activated T cell costimulatory receptor CD28.
    J Immunol. 2000 Jun 1;164(11):5805-14 PMID: 10820259
  15. SH3 domain recognition of a proline-independent tyrosine-based RKxxYxxY motif in immune cell adaptor SKAP55.
    EMBO J. 2000 Jun 15;19(12):2889-99 PMID: 10856234
  16. The adaptor protein Gads (Grb2-related adaptor downstream of Shc) is implicated in coupling hemopoietic progenitor kinase-1 to the activated TCR.
    J Immunol. 2000 Aug 1;165(3):1417-26 PMID: 10903746
  17. Identification of an atypical Grb2 carboxyl-terminal SH3 domain binding site in Gab docking proteins reveals Grb2-dependent and -independent recruitment of Gab1 to receptor tyrosine kinases.
    J Biol Chem. 2000 Oct 6;275(40):31536-45 PMID: 10913131
  18. Structure of a c-Cbl-UbcH7 complex: RING domain function in ubiquitin-protein ligases.
    Cell. 2000 Aug 18;102(4):533-9 PMID: 10966114
  19. A deubiquitinating enzyme UBPY interacts with the Src homology 3 domain of Hrs-binding protein via a novel binding motif PX(V/I)(D/N)RXXKP.
    J Biol Chem. 2000 Dec 1;275(48):37481-7 PMID: 10982817
  20. Lymphocytes with a complex: adapter proteins in antigen receptor signaling.
    Immunol Today. 2000 Nov;21(11):584-91 PMID: 11094263
  21. The peroxisomal membrane protein Pex13p shows a novel mode of SH3 interaction.
    EMBO J. 2000 Dec 1;19(23):6382-91 PMID: 11101511
  22. Requirement for the SLP-76 adaptor GADS in T cell development.
    Science. 2001 Mar 9;291(5510):1987-91 PMID: 11239162
  23. Ligand-independent assembly of recombinant human CD1 by using oxidative refolding chromatography.
    Proc Natl Acad Sci U S A. 2001 Mar 13;98(6):3288-93 PMID: 11248071
  24. SH3 domains: complexity in moderation.
    J Cell Sci. 2001 Apr;114(Pt 7):1253-63 PMID: 11256992
  25. The C-terminal SH3 domain of the adapter protein Grb2 binds with high affinity to sequences in Gab1 and SLP-76 which lack the SH3-typical P-x-x-P core motif.
    Oncogene. 2001 Mar 1;20(9):1052-62 PMID: 11314042
  26. Effect of pH and salt bridges on structural assembly: molecular structures of the monomer and intertwined dimer of the Eps8 SH3 domain.
    Protein Sci. 2001 May;10(5):1046-55 PMID: 11316885
  27. Novel recognition mode between Vav and Grb2 SH3 domains.
    EMBO J. 2001 Jun 15;20(12):2995-3007 PMID: 11406576
  28. Structural characteristics of protein binding sites for calcium and lanthanide ions.
    J Biol Inorg Chem. 2001 Jun;6(5-6):479-89 PMID: 11472012
  29. The role of Gads in hematopoietic cell signalling.
    Oncogene. 2001 Oct 1;20(44):6284-90 PMID: 11607830
  30. A novel, specific interaction involving the Csk SH3 domain and its natural ligand.
    Nat Struct Biol. 2001 Nov;8(11):998-1004 PMID: 11685249
  31. An Src homology 3-like domain is responsible for dimerization of the repressor protein KorB encoded by the promiscuous IncP plasmid RP4.
    J Biol Chem. 2002 Feb 8;277(6):4191-8 PMID: 11711548
  32. Maximum-likelihood density modification using pattern recognition of structural motifs.
    Acta Crystallogr D Biol Crystallogr. 2001 Dec;57(Pt 12):1755-62 PMID: 11717487
  33. Recent improvements to the SMART domain-based sequence annotation resource.
    Nucleic Acids Res. 2002 Jan 1;30(1):242-4 PMID: 11752305
  34. Solution structure of N-terminal SH3 domain of Vav and the recognition site for Grb2 C-terminal SH3 domain.
    J Biomol NMR. 2002 Jan;22(1):37-46 PMID: 11885979
  35. Scaffolds, adaptors and linkers of TCR signaling: theory and practice.
    Curr Opin Immunol. 2002 Jun;14(3):312-6 PMID: 11973128
  36. Induced expression and association of the Mona/Gads adapter and Gab3 scaffolding protein during monocyte/macrophage differentiation.
    Mol Cell Biol. 2002 Jun;22(11):3744-56 PMID: 11997510
  37. SH3 domain-mediated binding of the Drk protein to Dos is an important step in signaling of Drosophila receptor tyrosine kinases.
    Mech Dev. 2002 Aug;116(1-2):129-39 PMID: 12128212
  38. Diverse recognition of non-PxxP peptide ligands by the SH3 domains from p67(phox), Grb2 and Pex13p.
    EMBO J. 2002 Aug 15;21(16):4268-76 PMID: 12169629
  39. A high-affinity Arg-X-X-Lys SH3 binding motif confers specificity for the interaction between Gads and SLP-76 in T cell signaling.
    Curr Biol. 2002 Aug 6;12(15):1336-41 PMID: 12176364
  40. Structural determinants for the biological activity of Vav proteins.
    J Biol Chem. 2002 Nov 22;277(47):45377-92 PMID: 12228230
  41. How SH3 domains recognize proline.
    Adv Protein Chem. 2002;61:211-68 PMID: 12461825
  42. Expression of the Grb2-related protein of the lymphoid system in B cell subsets enhances B cell antigen receptor signaling through mitogen-activated protein kinase pathways.
    J Immunol. 2003 Jan 1;170(1):349-55 PMID: 12496419
  43. Raster3D Version 2.0. A program for photorealistic molecular graphics.
    Acta Crystallogr D Biol Crystallogr. 1994 Nov 1;50(Pt 6):869-73 PMID: 15299354
  44. Automated refinement of protein models.
    Acta Crystallogr D Biol Crystallogr. 1993 Jan 1;49(Pt 1):129-47 PMID: 15299554
  45. Refinement of macromolecular structures by the maximum-likelihood method.
    Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55 PMID: 15299926
  46. Protein folding and association: insights from the interfacial and thermodynamic properties of hydrocarbons.
    Proteins. 1991;11(4):281-96 PMID: 1758883
  47. Improved methods for building protein models in electron density maps and the location of errors in these models.
    Acta Crystallogr A. 1991 Mar 1;47 ( Pt 2):110-9 PMID: 2025413
  48. Processing of X-ray diffraction data collected in oscillation mode.
    Methods Enzymol. 1997;276:307-26 PMID: 27754618
  49. Multiple sequence alignment with hierarchical clustering.
    Nucleic Acids Res. 1988 Nov 25;16(22):10881-90 PMID: 2849754
  50. Crystal structure of cat muscle pyruvate kinase at a resolution of 2.6 A.
    J Mol Biol. 1979 Oct 15;134(1):109-42 PMID: 537059
  51. Dictionary of protein secondary structure: pattern recognition of hydrogen-bonded and geometrical features.
    Biopolymers. 1983 Dec;22(12):2577-637 PMID: 6667333
  52. Atomic structures of the human immunophilin FKBP-12 complexes with FK506 and rapamycin.
    J Mol Biol. 1993 Jan 5;229(1):105-24 PMID: 7678431
  53. Molecular cloning of SLP-76, a 76-kDa tyrosine phosphoprotein associated with Grb2 in T cells.
    J Biol Chem. 1995 Mar 31;270(13):7029-32 PMID: 7706237
  54. Structural basis for the specific interaction of lysine-containing proline-rich peptides with the N-terminal SH3 domain of c-Crk.
    Structure. 1995 Feb 15;3(2):215-26 PMID: 7735837
  55. 2,6-diacetylpyridine bis(thiosemicarbazones) zinc complexes: synthesis, structure, and biological activity.
    J Inorg Biochem. 1995 May 15;58(3):157-75 PMID: 7782786
  56. Structural determinants of peptide-binding orientation and of sequence specificity in SH3 domains.
    Nature. 1994 Nov 24;372(6504):375-9 PMID: 7802869
  57. Searching protein structure databases has come of age.
    Proteins. 1994 Jul;19(3):165-73 PMID: 7937731
  58. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice.
    Nucleic Acids Res. 1994 Nov 11;22(22):4673-80 PMID: 7984417
  59. Crystal structure of globular domain of histone H5 and its implications for nucleosome binding.
    Nature. 1993 Mar 18;362(6417):219-23 PMID: 8384699
  60. Reading between the lines: SH3 recognition of an intact protein.
    Structure. 1996 Jun 15;4(6):657-9 PMID: 8805558
  61. Cytoplasmic signalling domains: the next generation.
    Trends Biochem Sci. 1997 Aug;22(8):296-8 PMID: 9270302
  62. The SH3 domain of Eps8 exists as a novel intertwined dimer.
    Nat Struct Biol. 1997 Sep;4(9):739-43 PMID: 9303002
  63. An extensively modified version of MolScript that includes greatly enhanced coloring capabilities.
    J Mol Graph Model. 1997 Apr;15(2):132-4, 112-3 PMID: 9385560
  64. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.
    Nucleic Acids Res. 1997 Dec 15;25(24):4876-82 PMID: 9396791
  65. Amyloid fibril formation by an SH3 domain.
    Proc Natl Acad Sci U S A. 1998 Apr 14;95(8):4224-8 PMID: 9539718
  66. SMART, a simple modular architecture research tool: identification of signaling domains.
    Proc Natl Acad Sci U S A. 1998 May 26;95(11):5857-64 PMID: 9600884
  67. UBPY: a growth-regulated human ubiquitin isopeptidase.
    EMBO J. 1998 Jun 15;17(12):3241-50 PMID: 9628861
  68. PAK kinases are directly coupled to the PIX family of nucleotide exchange factors.
    Mol Cell. 1998 Jan;1(2):183-92 PMID: 9659915
  69. Crystallography & NMR system: A new software suite for macromolecular structure determination.
    Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905-21 PMID: 9757107
  70. The significance of zinc for leukocyte biology.
    J Leukoc Biol. 1998 Nov;64(5):571-7 PMID: 9823760
  71. Zinc is essential for binding of p56(lck) to CD4 and CD8alpha.
    J Biol Chem. 1998 Dec 4;273(49):32878-82 PMID: 9830036
  72. Mona, a novel hematopoietic-specific adaptor interacting with the macrophage colony-stimulating factor receptor, is implicated in monocyte/macrophage development.
    EMBO J. 1998 Dec 15;17(24):7273-81 PMID: 9857184
  73. Gads is a novel SH2 and SH3 domain-containing adaptor protein that binds to tyrosine-phosphorylated Shc.
    Oncogene. 1998 Dec 17;17(24):3073-82 PMID: 9872323
  74. Molecular cloning and expression of human grap-2, a novel leukocyte-specific SH2- and SH3-containing adaptor-like protein that binds to gab-1.
    Biochem Biophys Res Commun. 1998 Dec 18;253(2):443-7 PMID: 9878555
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/cdg258
Published
2003-06-02
Pages
2571-82
Language
English
Region
England
NLM ID
8208664
PMCID
PMC156755
Subset
IM
Grants
Wellcome Trust · United Kingdom
Databases
PDB
1OEB.PDB.
External Links
PubMed source DOI: 10.1093/emboj/cdg258 PMC Full Text View journal details
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