Skip to main content
SpringerLink
Log in

Chemical shift assignments and secondary structure of the Grb2 SH2 domain by heteronuclear NMR spectroscopy

  • Research Papers
  • Published:
Journal of Biomolecular NMR Aims and scope Submit manuscript

Summary

The growth factor receptor-bound protein-2 (Grb2) is an adaptor protein that mediates signal transduction pathways. Chemical shift assignments were obtained for the SH2 domain of Grb2 by heteronuclear NMR spectroscopy, employing the uniformly 13C-/15N-enriched protein as well as the protein containing selectively 15N-enriched amino acids. Using the Chemical Shift Index (CSI) method, the chemical shift indices of four nuclei, 1Hα, 13Cα, 13Cβ and 13CO, were used to derive the secondary structure of the protein. Nuclear Overhauser enhancements (NOEs) were then employed to confirm the secondary structure. The CSI results were compared to the secondary structural elements predicted for the Grb2 SH2 domain from a sequence alignment [Lee et al. (1994) Structure, 2, 423–438]. The core structure of the SH2 domain contains an antiparallel β-sheet and two α-helices. In general, the secondary structural elements determined from the CSI method agree well with those predicted from the sequence alignment.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

crk:

viral p47gag-crk

EGF:

epidermal growth factor

GAP:

GTPase-activating protein

PI3K:

phosphatidylinositol-3-kinase

PLC-γ:

phospholipase-C-γ, shc, src homologous and collagen

src:

sarcoma family of nonreceptor tyrosine kinase

References

  • Bax, A., Clore, G.M. and Gronenborn, A.M. (1990) J. Magn. Reson., 88, 425–431.

    Google Scholar 

  • Bax, A. and Ikura, M. (1991) J. Biomol. NMR, 1, 99–104.

    Google Scholar 

  • Bodenhausen, G. and Ruben, D.J. (1980) Chem. Phys. Lett., 69, 185–189.

    Google Scholar 

  • Booker, G.W., Breeze, A.L., Downing, A.K., Panayotou, G., Gout, I., Waterfield, M.D. and Campbell, I.D. (1992) Nature, 358, 684–687.

    Google Scholar 

  • Brown, S.C., Weber, P.L. and Mueller, L. (1988) J. Magn. Reson., 77, 166–169.

    Google Scholar 

  • Chardin, P., Camonis, J.H., Gale, N.W., Van, Aelst, L., Schlessinger, J., Wigler, M.H. and Bar-Sagi, D. (1993) Science, 260, 1338–1343.

    Google Scholar 

  • Clark, S.G., Stern, M.J. and Horvitz, H.R. (1992) Nature, 356, 340–344.

    Google Scholar 

  • DeLoskey, R.J., Van, Dyk, D.E., Van, Aken, T.E. and Campbell-Burk, S. (1994) Arch. Biochem. Biophys., 311, 72–78.

    Google Scholar 

  • Downward, J. (1994) FEBS Lett., 338, 113–117.

    Google Scholar 

  • Eck, M.J., Shoelson, S.E. and Harrison, S.C. (1993) Nature, 362, 87–91.

    Google Scholar 

  • FarmerII, B.T., Venters, R.A., Spicer, L.D., Wittekind, M.G. and Mueller, L. (1992) J. Biomol. NMR, 2, 195–202.

    Google Scholar 

  • Griesinger, C., Otting, G., Wüthrich, K. and Ernst, R.R. (1988) J. Am. Chem. Soc., 110, 7870–7872.

    Google Scholar 

  • Grzesiek, S. and Bax, A. (1992) J. Am. Chem. Soc., 114, 6291–6293.

    Google Scholar 

  • Hatada, M.H., Lu, X., Laird, E.R., Green, J., Morgenstern, J.P., Lou, M., Marr, C.S., Philips, T.B., Ram, M.K., Theriault, K., Zoller, M.J. and Karas, J.L. (1995) Nature, 377, 32–38.

    Google Scholar 

  • Hensmann, M., Booker, G.W., Panayotou, G., Boyd, J., Linacre, J., Waterfield, M. and Campbell, I.D. (1994) Protein Sci., 3, 1020–1030.

    Google Scholar 

  • Kay, L.E., Ikura, M., Tschudin, R. and Bax, A. (1990) J. Magn. Reson., 89, 496–514.

    Google Scholar 

  • Kay, L.E., Xu, G.Y., Singer, A.U., Muhandiram, D.R. and Forman-Kay, J.D. (1993) J. Magn. Reson. Ser. B, 101, 333–337.

    Google Scholar 

  • Lee, C.H., Kominos, D., Jacques, S., Margolis, B., Schlessinger, J., Shoelson, S.E. and Kuriyan, J. (1994) Structure, 2, 423–438.

    Google Scholar 

  • LeMaster, D.M. and Richards, F.M. (1988) Biochemistry, 27, 142–150.

    Google Scholar 

  • Levy, G.C. and Lichter, R.L. (Eds) (1979) Nitrogen-15 Nuclear Magnetic Resonance Spectroscopy, Wiley, New York, NY.

    Google Scholar 

  • Lowenstein, E.J., Daly, R.J., Batzer, A.G., Li, W., Margolis, B., Lammers, R., Ullrich, A., Skolnik, E.Y., Bar-Sagi, D. and Schlessinger, J. (1992) Cell, 70, 431–442.

    Google Scholar 

  • Maignan, S., Guilloteau, J.P., Fromage, N., Arnoux, B., Becquart, J. and Ducruix, A. (1995) Science, 268, 291–293.

    Google Scholar 

  • Marion, D. and Wüthrich, K. (1983) Biochem. Biophys. Res. Commun., 113, 967–974.

    Google Scholar 

  • Marion, D., Ikura, M., Tschudin, R. and Bax, A. (1989a) J. Magn. Reson., 85, 393–399.

    Google Scholar 

  • Marion, D., Driscoll, P.C., Kay, L.E., Wingfield, P.T., Bax, A., Gronenborn, A.M. and Clore, G.M. (1989b) Biochemistry, 28, 6150–6156.

    Google Scholar 

  • McIntosh, L.P. and Dahlquist, F.W. (1990) Q. Rev. Biophys., 23, 1–38.

    Google Scholar 

  • Muchmore, D.C., McIntosh, L.P., Russell, C.B., Anderson, D.E. and Dahlquist, F.W. (1989) Methods Enzymol., 177, 44–73.

    Google Scholar 

  • Muhandiram, D.R. and Kay, L.E. (1994) J. Magn. Reson. Ser. B, 103, 203–216.

    Google Scholar 

  • Narula, S.S., Yuan, R.W., Adams, S.E., Green, O.M., Green, J., Philips, T.B., Zydowsky, L.D., Botfield, M.C., Hatada, M., Laird, E.R., Zoller, M.J., Karas, J.L. and Dalgarno, D.C. (1995) Structure, 3, 1061–1073.

    Google Scholar 

  • Olivier, J.P., Raabe, T., Henkemeyer, M., Dickson, B., Mbamalu, G., Margolis, B., Schlessinger, J., Hafen, E. and Pawson, T. (1993) Cell, 73, 179–191.

    Google Scholar 

  • Overduin, M., Mayer, B., Rios, C.B., Baltimore, D. and Cowburn, D. (1992a) Proc. Natl. Acad. Sci. USA, 89, 11673–11677.

    Google Scholar 

  • Overduin, M., Rios, C.B., Mayer, B.J., Baltimore, D. and Cowburn, D. (1992b) Cell, 70, 697–704.

    Google Scholar 

  • Pascal, S.M., Singer, A.U., Gish, G., Yamazaki, T., Shoelson, S.E., Pawson, T., Kay, L.E. and Forman-Kay, J.D. (1994) Cell, 77, 461–472.

    Google Scholar 

  • Pawson, T. (1995) Nature, 373, 573–580.

    Google Scholar 

  • Shaka, A.J., Keeler, J. and Freeman, R. (1983) J. Magn. Reson., 53, 313–340.

    Google Scholar 

  • Shaka, A.J., Barker, P.B. and Freeman, R. (1985) J. Magn. Reson., 64, 547–552.

    Google Scholar 

  • Shaka, A.J., Lee, C.J. and Pines, A. (1988) J. Magn. Reson., 77, 274–293.

    Google Scholar 

  • Songyang, Z., Shoelson, S.E., McGlade, J., Olivier, P., Pawson, T., Bustelo, X.R., Barbacid, M., Sabe, H., Hanafusa, H., Yi, T., Ren, R., Baltimore, D., Ratnofsky, S., Feldman, R.A. and Cantley, L.C. (1994) Mol. Cell. Biol., 14, 2777–2785.

    Google Scholar 

  • States, D.J., Haberkorn, R.A. and Ruben, D.J. (1982) J. Magn. Reson., 48, 286–292.

    Google Scholar 

  • Waksman, G., Kominos, D., Robertson, S.C., Pant, N., Baltimore, G., Birge, R.B., Cowburn, D., Hanafusa, H., Mayer, B.J., Overduin, M., Resh, M.D., Rios, C.B., Silverman, L. and Kuriyan, J. (1992) Nature, 358, 646–653.

    Google Scholar 

  • Waksman, G., Shoelson, S.E., Pant, N., Cowburn, D. and Kuriyan, J. (1993) Cell, 72, 779–790.

    Google Scholar 

  • Wishart, D.S. and Sykes, B.D. (1994) J. Biomol. NMR, 4, 171–180.

    Google Scholar 

  • Wittekind, M. and Mueller, L. (1993) J. Magn. Reson. Ser. B, 101, 201–205.

    Google Scholar 

  • Xu, R.X., Word, J.M., Davis, D.G., Rink, M.J., Willard, D.H. and GampeJr., R.T. (1995) Biochemistry 34, 2107–2121.

    Google Scholar 

  • Zhou, M.M., Meadows, R.P., Logan, T.M., Yoon, H.S., Wade, W.S., Ravichandran, K.S., Burakoff, S.J. and Fesik, S.W. (1995) Proc. Natl. Acad. Sci. USA, 92, 7784–7788.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Schering-Plough Research Institute, 2015 Galloping Hill Road, 07033, Kenilworth, NJ, USA

    Yu-Sen Wang, Anne F. Frederick, Mary M. Senior, Barbara A. Lyons, Stuart Black, Paul Kirschmeier, Louise M. Perkins & Oswald Wilson

  2. Department of Biochemistry, College of Medicine, University of Vermont, 05405, Burlington, VT, USA

    Barbara A. Lyons

Authors
  1. Yu-Sen Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anne F. Frederick
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mary M. Senior
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Barbara A. Lyons
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Stuart Black
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Paul Kirschmeier
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Louise M. Perkins
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Oswald Wilson
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wang, YS., Frederick, A.F., Senior, M.M. et al. Chemical shift assignments and secondary structure of the Grb2 SH2 domain by heteronuclear NMR spectroscopy. J Biomol NMR 7, 89–98 (1996). https://doi.org/10.1007/BF00203819

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00203819

Keywords

Search

Navigation