Article | Published:

The SH3 domain of Eps8 exists as a novel intertwined dimer

Nature Structural Biologyvolume 4pages739743 (1997) | Download Citation



SH3 domains are structurally well-characterized as monomeric modular units of protein structure that mediate protein–protein recognition in numerous signal transduction proteins. The X-ray crystallographic structure of the Eps8 SH3 domain reveals a novel variation of the canonical SH3 fold: the SH3 domain from Eps8 is a dimer formed by strand interchange. In addition, co-immunoprecipitation experiments show that intact Eps8 is multimeric in vivo. Hence, the SH3 domain of Eps8 may represent a dimerization motif.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1

    Morton, C.J. et al., Solution structure and peptide binding of the SH3 domain from human Fyn. Structure 4, 705–714 (1996).

  2. 2

    Yu, H. et al., Structural basis for the binding of proline rich peptides to SH3 domains. Cell 76, 933–945 (1994).

  3. 3

    Saraste, M. and Musacchio, A. Backwards and forward binding. Nature Struct. Biol. 1, 835–837 (1994).

  4. 4

    Feng, S., Chen, J.K., Yu, H., Simon, J.A. and Schreiber, S.L. Two binding orientations for peptides to the Src SH3 domain: development of a general model for SH3-ligand interaction. Science 266, 1241–1247 (1994).

  5. 5

    Musacchio, A., Saraste, M. and Wilmanns, M. High resolution crystal structures of tyrosine kinase SH3 domains complexed with proline rich peptides. Nature Struct. Biol. 1, 546–551 (1994).

  6. 6

    Lim, W.A., Richards, F.M. and Fox, R.O. Structural determinants of peptide-binding orientation and of sequence specificity on SH3 domains. Nature 372, 375–379 (1994).

  7. 7

    Terasawa, H. et al., Structure of the N-terminal SH3 domain of GRB2 complexed with a peptide from the guanine nucleotide releasing factor Sos. Nature Struct. Biol. 1, 891–897 (1994).

  8. 8

    Goudreau, N. et al., NMR structure of the N-terminal SH3 domain of GRB2 and its complex with a praline-rich peptide from SOS. Nature Struct. Biol. 1, 898–907 (1994).

  9. 9

    Musacchio, A., Noble, M.E.M., Pauptil, R., Wierenga, R.K. and Saraste, M. Crystal structure of a Src-homology 3 (SH3) domain. Nature 359, 851–855 (1992).

  10. 10

    Noble, M.E.M., Musachio, A., Saraste, M., Courtneidge, S.A. and Wierenga, R.K. Crystal structure of the SH3 domain in human Fyn: a comparison of the three-dimensional structures of SH3 domains in tyrosine kinases and spectrin. EMBO J. 12, 2617–2624 (1993).

  11. 11

    Booker, G.W. et al., Solution structure and ligand binding site of the SH3 domain of the p85 alpha subunit of phosphoinositol 3-kinase. Cell 73, 813–822 (1993).

  12. 12

    Yang, Y.S. et al., Solution structure of GAP SH3 domain by 1H NMR and spatial arrangement of essential Ras signaling-involved sequence. EMBO J. 72, 1270–1279 (1994).

  13. 13

    Kohda, D. et al., Solution structure of the SH3 domain of phospholipase-c gamma. Cell 72, 953–960 (1993).

  14. 14

    Borchert, T.V., Mathieu, M., Zeelen, J.P., Courtneidge, S.A. and Wierenga, R.K. The crystal structure of human CskSH3: structural diversity near the RT-src and n-src loop. FEBS Lett. 341, 79–85 (1994).

  15. 15

    Kohda, D. et al., Solution structure and ligand binding site of the carboxyterminal SH3 domain of GRB2. Structure 2, 1029–1040 (1994)

  16. 16

    Fazioli, F. et al., Eps8, a substrate for the epidermal growth factor receptor kinase, enhances EGF-dependent mitogenic signals. EMBO J. 12, 3799–3808 (1993).

  17. 17

    Fazioli, F., Minichiello, L, Matoskova, B., Wong, W.T. and DiFiore, P.P. Eps15, a novel tyrosine kinase substrate exhibits transforming activity. Mol. Cell Biol. 13, 5814–5828 (1993).

  18. 18

    Castagnino, P., Biesova, Z., Wong, W.T., Fazioli, F., Gill, G. and DiFiore, P.P. Direct binding of Eps8 to the juxtamembrane domain of EGFR is phosphotyrosine and SH2 independent. Oncogene 10, 723–729 (1995).

  19. 19

    Matoskova, B., Wong, W.T., Nomura, N., Robbins, K.C. and Di Fiore, P.P. RN-tre specifically binds to the SH3 domain of Eps8 with high affinity and confers growth advantage to NIH3T3 upon carboxy-terminal truncation. Oncogene 12, 2679–2688 (1996).

  20. 20

    Bennet, M.J., Schlunegger, M.P. and Eisenberg, D. 3D domain swapping: a mechanism for oligomer assembly. Prot. Sci. 4, 2455–2468 (1995).

  21. 21

    Y., Yan, et al., Crystal structure of repetitive segments of spectrin. Science 262, 2027–2030 (1993).

  22. 22

    Parge, H.E., Arvai, A.S., Mustari, D.J., Reed, S.I. and Tainer, J.A. Human CksHs2 atomic structure: a role for its hexameric assembly in cell cycle control. Science 262, 387–395 (1993).

  23. 23

    Hunter, C.A., Singh, J. and Thornton, J.M. Pi-Pi interactions: the geometry and energetics of phenylalanine-phenylalanine interactions in proteins. J. Mol. Biol. 218, 837–846 (1991).

  24. 24

    Wu, X. et al., Structural basis for the specific interaction of lysine-containing proline-rich peptides with the N-terminal SH3 domain of c-CRK. Structure 3, 215–226 (1995).

  25. 25

    Hatada, M.H. et al., Molecular basis for interaction of the protein tyrosine kinase ZAP-70 with the T-cell receptor. Nature 377, 835–837 (1995).

  26. 26

    Zhang, O. and Forman-Kay, J.D. Structural characterization of folded and unfolded states of and SH3 domain in equilibrium in aqueous buffer. Biochemistry 34, 6784–6792 (1995).

  27. 27

    Matoskova, B., Wong, W.T., Salcini, A.E., Pelicci, P.G. and Di Fiore, P.P. Constitutive phosphosphorylation of Eps8 in tumor cell lines: relevance to malignant transformation. Mol. Cell. Biol. 15, 3805–3812 (1995).

  28. 28

    Guan, K.L. and Dixon, J.E. Eukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure of fusion proteins with glutathione-S-transferase. Anal. Biochem. 192, 262–267 (1991).

  29. 29

    Otwinowski, Z. Proceedings of the CCP4 Study Weekend: Data Collection and Processing (Eds. L. Sawyer, N. Isaacs, S. Bailey), SERC Daresbury Laboratory, Warrington, UK, 56 (1993).

  30. 30

    The CCP4 Suite: Programs for Protein Crystallography. Acta Crystallogr. D50, 760 (1994).

  31. 31

    Brünger, AT. X-PLOR, Version 3.1: A System for X-ray Crystallography and NMR. Yale University Press, New Haven, CT.

  32. 32

    Tronrud, D.E. Conjugate-direction minimization: an improved method for the refinement of macromolecules. Acta Crystallogr. A48, 912–916 (1992).

  33. 33

    Brünger, AT. The free R value: a novel statistical quantity for assessing the accuracy of crystal structures. Nature 335, 472–474 (1992).

  34. 34

    Jones, T.A., Zou, J.Y., Cowan, S.W. and Kjeldgaard, M. Improved methods for building protein models in electron density maps and location of errors in these models. Acta Gyst. A47, 110–119 (1991).

  35. 35

    Read, R.J. Improved Fourier coefficients for maps using phases from partial structures with errors. Acta Crystallogr. A42, 140–149 (1986).

Download references

Author information


  1. Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, 37232-0146, USA

    • K.V. Radha Kishan
  2. Department of Experimental Oncology, European Institute of Oncology, Milan, Italy

    • Giorgio Scita
  3. Laboratory of Cellular Development and Oncology, National Institute of Dental Research, National Institutes of Health, Bethesda, MD, 20892, USA

    • William T. Wong
  4. Department of Experimental Oncology, European Institute of Oncology, Milan, Italy

    • Pier Paolo Di Fiore
  5. Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, 37232-0146, USA

    • Marcia E. Newcomer
  6. Istituto di Microbiologia, Facolta di Medicina e Chirurgia, Bari, Italy

    • Pier Paolo Di Fiore
    •  & Marcia E. Newcomer


  1. Search for K.V. Radha Kishan in:

  2. Search for Giorgio Scita in:

  3. Search for William T. Wong in:

  4. Search for Pier Paolo Di Fiore in:

  5. Search for Marcia E. Newcomer in:

About this article

Publication history



Issue Date


Further reading