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Solution structure of the Grb2 SH2 domain complexed with a high-affinity inhibitor

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Abstract

The solution structure of the growth factor receptor-bound protein 2 (Grb2) SH2 domain complexed with a high-affinity inhibitor containing a non-phosphorus phosphate mimetic within a macrocyclic platform was determined by nuclear magnetic resonance (NMR) spectroscopy. Unambiguous assignments of the bound inhibitor and intermolecular NOEs between the Grb2 SH2 domain and the inhibitor was accomplished using perdeuterated Grb2 SH2 protein. The well-defined solution structure of the complex was obtained and compared to those by X-ray crystallography. Since the crystal structure of the Grb2 SH2 domain formed a domain-swapped dimer and several inhibitors were bound to a hinge region, there were appreciable differences between the solution and crystal structures. Based on the binding interactions between the inhibitor and the Grb2 SH2 domain in solution, we proposed a design of second-generation inhibitors that could be expected to have higher affinity.

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Acknowledgement

This work was supported by Grant-in-Aid for Scientific Research and the National Projects on Protein Structure and Functional Analysis from the Ministry of Education, Science and Culture of Japan. This research was also supported in part by the International Research Program of the NIH, Center for Cancer Research, National Cancer Institute.

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Authors and Affiliations

  1. Department of Structural Biology, Graduate School of Pharmaceutical Sciences, Hokkaido University, N12 W6, Kita-ku, Sapporo, 060-0812, Japan

    Kenji Ogura, Takanori Shiga, Masashi Yokochi, Satoru Yuzawa & Fuyuhiko Inagaki

  2. National Cancer Institute at Frederick, Laboratory of Medicinal Chemistry, Center for Cancer Research, P. O. Box B, Frederick, MD, 21702-1201, USA

    Terrence R. Burke Jr.

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  1. Kenji Ogura
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  2. Takanori Shiga
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  3. Masashi Yokochi
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  4. Satoru Yuzawa
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  5. Terrence R. Burke Jr.
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  6. Fuyuhiko Inagaki
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Correspondence to Fuyuhiko Inagaki.

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Ogura, K., Shiga, T., Yokochi, M. et al. Solution structure of the Grb2 SH2 domain complexed with a high-affinity inhibitor. J Biomol NMR 42, 197–207 (2008). https://doi.org/10.1007/s10858-008-9272-0

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