title: Preparation of crystals for characterizing the Grb7 SH2 domain before and after complex formation with a bicyclic peptide antagonist.
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.
authors.affiliation: Department of Biochemistry and Molecular Biology, Monash University, VIC 3800, Australia.
authors.forename: Nigus D
authors.initials: ND
authors.lastname: Ambaye
authors.affiliation: Department of Biochemistry and Molecular Biology, Monash University, VIC 3800, Australia.
authors.forename: Menachem J
authors.initials: MJ
authors.lastname: Gunzburg
authors.affiliation: Department of Biochemistry and Molecular Biology, Monash University, VIC 3800, Australia.
authors.forename: Daouda A K
authors.initials: DA
authors.lastname: Traore
authors.affiliation: School of Chemistry, Monash University, VIC 3800, Australia.
authors.forename: Mark P
authors.initials: MP
authors.lastname: Del Borgo
authors.affiliation: School of Chemistry, Monash University, VIC 3800, Australia.
authors.forename: Patrick
authors.initials: P
authors.lastname: Perlmutter
authors.affiliation: Department of Biochemistry and Molecular Biology, Monash University, VIC 3800, Australia.
authors.forename: Matthew C J
authors.initials: MC
authors.lastname: Wilce
authors.affiliation: Department of Biochemistry and Molecular Biology, Monash University, VIC 3800, Australia.
authors.forename: Jacqueline A
authors.initials: JA
authors.lastname: Wilce
mesh_terms: D000595:Amino Acid Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray; D051382:GRB7 Adaptor Protein / Q000737:chemistry*; D006801:Humans; D010456:Peptides, Cyclic / Q000737:chemistry*; D011487:Protein Conformation; D018909:src Homology Domains
chemical_list: C495955:GRB7 protein, human; D010456:Peptides, Cyclic; D051382:GRB7 Adaptor Protein
keywords: Grb7; SH2 domain; seeding