Abstract
GRB2 is a small adaptor protein of 217 amino acids comprising one SH2 domain surrounded by two SH3 domains. GRB2 couples receptor tyrosine kinase activation to Ras signalling by interacting, through its SH3 domains, to the carboxy-terminal proline-rich regions of the guanine nucleotide exchange factor Sos. Here we report the synthesis and solution structure of the amino-terminal SH3 domain of GRB2 and of its more stable Ser 32 mutant. 1H NMR analysis of the complex between the Ser-32-GRB2-N-SH3 domain and the proline-rich peptide VPPPVPPRRR, derived from h-Sos, shows that relative to the SH3 peptide complexes described for PI3K, Fyn and Abl, the proline-rich peptide in this complex binds in the opposite orientation.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$189.00 per year
only $15.75 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Ullrich, A. & Schlessinger, J. Signal transduction by receptors with tyrosine kinase activity. Cell 61, 203–212 (1990).
Cantley, L.C. et al. Oncogenes and signal transduction. Cell 64, 281–302 (1991).
Fry, M.J., Panayotou, G., Booker, G.W. & Waterfield, M.D. New insights into protein-tyrosine kinase receptor signaling complexes. Prot. Sci. 2, 1785–1797 (1993).
Pawson, T. & Gish, G.D. SH2 and SH3 domains: from structure to function. Cell 71, 359–362 (1992).
Pawson, T. & Schlessinger, J. SH2 and SH3 domains. Curr. Biol. 3, 434–442 (1993).
Koch, C.A., Anderson, D., Moran, M.F., Ellis, C. & Pawson, T. SH2 and SH3 domains: elements that control interactions of cytoplasmic signaling proteins. Science 252, 668–674 (1991).
Musacchio, A., Gibson, T., Lehto, V.P. & Saraste, M. SH3 - an abundant protein domain in search of a function. FEBS Letts. 307, 55–61 (1992).
Lowenstein, E.J. et al. The SH2 and SH3 domain-containing protein Grb2 links receptor tyrosine kinases to ras signaling. Cell 70, 431–442 (1992).
Matuoka, K., Shibasaki, F., Shibata, M. & Takenawa, T. Ash/Grb-2, a SH2/SH3-containing protein, couples to signaling for mitogenesis and cytoskeletal reorganization by EGF AND PDGF EMBO J. 12, 3467–3473 (1993).
Skolnik, E.Y. et al. The SH2/SH3 domain-containing protein Grb2 interacts with tyrosine-phosphorylated IRS1 and Shc: Implications for insulin control of ras signaling. EMBO J. 12, 1929–1936 (1993).
Egan, S.E. et al. Association of Sos Ras exchange protein with Grb2 is implicated in tyrosine kinase sinal transduction and transformation. Nature 363, 45–51 (1993).
Chardin, P. et al. Human Sos 1: A guanine nucleotide exchange factor for Ras that binds Grb2. Science 260, 1338–1343 (1993).
Rozakis-Adcock, M., Fernley, R., Wade, J., Pawson, T. & Bowtell, D. The SH2 and SH3 domains of mammalian Grb2 couple the EGF receptor to Ras activator mSos1. Nature 363, 83–85 (1993).
Li, N. et al. Guanine-nucleotide releasing factor hSos1 binds to Grb2 and links receptor tyrosine kinases to Ras signaling. Nature 363, 85–88 (1993).
Yu, H. et al. Solution structure of the SH3 domain of src and identification of its ligand-binding site. Science 258, 1665–1668 (1992).
Koyama, S. et al. Structure of the PI3K SH3 domain and analysis of the SH3 family. Cell 72, 945–952 (1993).
Booker, G.W. et al. Solution structure and ligand-binding site of the SH3 domain of the p85a subunit of phosphatidylinositol 3-kinase. Cell 73, 813–822 (1993).
Kohda, D. et al. Solution structure of the SH3 domain of phospholipase C-γ. Cell 72, 953–960 (1993).
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. 13, 1270–1279 (1994).
Musacchio, A., Noble, M., Pauptit, R., Wierenga, R. & Saraste, M. Crystal structure of a Src-homology 3 (SH3) domain. Nature 359, 851–855 (1992).
Noble, M., Musacchio, A., Saraste, M., Courtneidge, S.A. & Wierenga, R. Crystal structure of the SH3 domain in human Fyn:comparison of the three-dimensional structures of SH3 domains in tyrosine kinases and spectrin. EMBO J. 12, 2617–2624 (1993).
Borchert, T.V., Mathieu, M., Zeelen, J.P., Courtneidge, S.A. & Wierenga, R.K. The crystal structure of human CskSH3: structural diversity near the RT-Src and n-Src loop. FEBS letts 341, 79–85 (1994).
Eck, M.J., Atwell, S.K., Shoelson, S.E. & Harrison, S.C. Structure of the regulatory domains of the Src-family tyrosine kinase Lck. Nature 368, 764–769 (1994).
Yu, H. et al. Structural basis for the binding of proline-rich peptides to SH3 domains. Cell 76, 933–945 (1994).
Musacchio, A., Saraste, M. & Wilmanns, M. High-resolution crystal structures of tyrosine kinase SH3 domains complexed with proline-rich peptides. Nature struct. Biol. 1, 546–551 (1994).
Nilges, M., Clore, G.M. & Gronenborn, A.M. Determination of three-dimensional structures of proteins from interproton distance data by dynamical simulated annealing from a random array or atoms. FEBS Letts 239, 129–134 (1988).
Olivier, J.P. et al. A drosophila SH2-SH3 adaptor protein implicated in coupling the sevenless tyrosine kinase to an activator of Ras guanine nucleotide exchange, Sos. 73, 179–191 (1993).
Lim, W.A. & Richards, F.M. Critical residues in an SH3 domain from Sem-5 suggest a machanism for praline-rich peptide recognition. Nature struct. Biol. 1, 221–225 (1994).
Trahey, M. et al. Molecular cloning of two types of GAP complementary DNA from human placenta. Science 242, 1697–1700 (1988).
Chen, J.K., Lane, W.S., Brauer, A.W., Tanaka, A. & Schreiber, S.L. Biased combinatorial libraries: novel ligands for the SH3 domain of phosphatidylinositol 3-kinase. J. Am. chem. Soc. 115, 12591–12592 (1993).
Cussac, D., Frech, M., & Chardin, P. Binding of the Grb2 SH2 domain to phosphotyrosine motifs does not change the affinity of its SH3 domains for Sos proline-rich motifs. EMBO J. 13, 4011–4021 (1994).
Wüthrich, K. NMR of proteins and nucleic acids. (Wileq, NewYork, N.Y.; 1986).
Rance, M. et al. Improved spectral resolution in COSY 1H NMR spectra of proteins via double quantum filtering. Biochem. biophys. Res. Commun. 117, 479–485 (1983).
Griesinger, C., Otting, G., Wuthrich, K. & Ernst, R.R. Clean TOCSY for 1H spin system identification in macromolecules. J. Am. chem. Soc. 110, 7870–7872 (1988).
Jeener, J., Meier, B.H., Bachmann, P. & Ernst, R.R. Investigation of exchange processes by two-dimensional NMR spectroscopy. J. chem. Phys. 71, 4546–4553 (1979).
Macura, S., Huang, Y., Suter, D. & Ernst, R.R. Two-dimensional chemical exchange and cross-relaxation spectroscopy of coupled nuclear-spins. J. magn. Reson. 43, 259–281 (1981).
Bax, A. & Davis, D.G. Pratical aspects of two-dimensional transverse NOE spectroscopy. J. magn. Reson. 63, 207–213 (1985).
Marion, D. & Wüthrich, K. Application of phase sensitive two-dimensional correlated spectroscopy (COSY) for measurements of 1H-1H spin-spin coupling constants in proteins. Biochem. biophys. Res. Commun. 113, 967–974 (1983).
Clore, G.M., Brünger, A.T., Karplus, M. & Gronenborn, A.M. Application of molecular dynamics with interproton distance restraints to three-dimensional protein structure determination. J. molec. Biol. 191, 523–551 (1986).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Goudreau, N., Cornille, F., Duchesne, M. et al. NMR structure of the N-terminal SH3 domain of GRB2 and its complex with a proline-rich peptide from Sos. Nat Struct Mol Biol 1, 898–907 (1994). https://doi.org/10.1038/nsb1294-898
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/nsb1294-898
This article is cited by
-
\(^1\)H, \(^{13}\)C and \(^{15}\)N assignments of human Grb2 free of ligands
Biomolecular NMR Assignments (2020)
-
Molecular Dynamics model of peptide-protein conjugation: case study of covalent complex between Sos1 peptide and N-terminal SH3 domain from Grb2
Scientific Reports (2019)
-
Conformational change of Sos-derived proline-rich peptide upon binding Grb2 N-terminal SH3 domain probed by NMR
Scientific Reports (2013)
-
Oligomerization of signaling complexes by the multipoint binding of GRB2 to both LAT and SOS1
Nature Structural & Molecular Biology (2006)
-
MET meet adaptors: Functional and structural implications in downstream signalling mediated by the Met receptor
Molecular and Cellular Biochemistry (2005)