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

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 groupP63, whereas the G7-B1–Grb7 SH2 domain complex crystallized in the monoclinic space groupP21. The experimental aspects of crystallization, crystal optimization and data collection and the preliminary data are reported.

authors

  • Ambaye, Nigus D
  • Gunzburg, Menachem J
  • Traore, Daouda AK
  • Del Borgo, Mark P
  • Perlmutter, Patrick
  • Wilce, Matthew CJ
  • Wilce, Jacqueline A

publication date

  • 2014