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. 2014 Feb;70(Pt 2):182-6.
doi: 10.1107/S2053230X13033414. Epub 2014 Jan 21.

Preparation of crystals for characterizing the Grb7 SH2 domain before and after complex formation with a bicyclic peptide antagonist

Nigus D Ambaye  1 , Menachem J Gunzburg  1 , Daouda A K Traore  1 , Mark P Del Borgo  2 , Patrick Perlmutter  2 , Matthew C J Wilce  1 , Jacqueline A Wilce  1
Affiliations

Preparation of crystals for characterizing the Grb7 SH2 domain before and after complex formation with a bicyclic peptide antagonist

Nigus D Ambaye et al. Acta Crystallogr F Struct Biol Commun. 2014 Feb.

Abstract

Human growth factor receptor-bound protein 7 (Grb7) is an adapter protein involved in cell growth, migration and proliferation. It is now recognized that Grb7 is an emerging therapeutic target in specific cancer subtypes. Recently, the discovery of a bicyclic peptide inhibitor that targets the Grb7 SH2 domain, named G7-B1, was reported. In an attempt to probe the foundation of its interaction with Grb7, the crystallization and preliminary data collection of both the apo and G7-B1-bound forms of the Grb7 SH2 domain are reported here. Diffraction-quality crystals were obtained using the hanging-drop vapour-diffusion method. After several rounds of microseeding, crystals of the apo Grb7 SH2 domain were obtained that diffracted to 1.8 Å resolution, while those of the G7-B1-Grb7 SH2 domain complex diffracted to 2.2 Å resolution. The apo Grb7 SH2 domain crystallized in the trigonal space group P63, whereas the G7-B1-Grb7 SH2 domain complex crystallized in the monoclinic space group P21. The experimental aspects of crystallization, crystal optimization and data collection and the preliminary data are reported.

Keywords: Grb7; SH2 domain; seeding.

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Figures

Figure 1
Figure 1
Schematic and full chemical structure of the G7-B1 peptide antagonist. Single-letter amino-acid abbreviations are indicated in blue.
Figure 2
Figure 2
The initial crystals of the apo Grb7 SH2 domain obtained from random screening. The crystal was found to diffract to 2.0 Å resolution.
Figure 3
Figure 3
The process of apo Grb7 SH2 crystal optimization by microseeding.
Figure 4
Figure 4
Monoclinic crystals of the G7-B1–Grb7 SH2 domain complex. Diffraction-quality crystals were obtained after microseeding.
Figure 5
Figure 5
Diffraction images of Grb7 SH2 domain crystals. (a) Diffraction image of the trigonal crystals of the apo Grb7 SH2 domain; (b) diffraction image of the G7-B1–Grb7 SH2 domain co-crystal.
See this image and copyright information in PMC

References

    1. Ambaye, N. D., Gunzburg, M. J., Lim, R. C. C., Price, J. T., Wilce, M. C. J. & Wilce, J. A. (2011). Bioorg. Med. Chem. 19, 693–701. - PubMed
    1. Ambaye, N. D., Pero, S. C., Gunzburg, M. J., Yap, M.-Y., Clayton, D. J., Del Borgo, M. P., Perlmutter, P., Aguilar, M.-I., Shukla, G. S., Peletskaya, E., Cookson, M. M., Krag, D. N., Wilce, M. C. J. & Wilce, J. A. (2011). J. Mol. Biol. 412, 397–411. - PubMed
    1. Daly, R. J. (1998). Cell. Signal. 10, 613–618. - PubMed
    1. Evans, P. (2006). Acta Cryst. D62, 72–82. - PubMed
    1. Giricz, O., Calvo, V., Pero, S. C., Krag, D. N., Sparano, J. A. & Kenny, P. A. (2012). Breast Cancer Res. Treat. 133, 607–615. - PubMed

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