Grb7-SH2 domain dimerisation is affected by a single point mutation

Corrine Joy Porter, Matthew CJ Wilce, Joel P Mackay, P J Leedman, Jacqueline Anne Wilce

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23 Citations (Scopus)

Abstract

Growth factor receptor bound protein 7 (Grb7) is an adaptor protein that is co-overexpressed and forms a tight complex with the ErbB2 receptor in a number of breast tumours and breast cancer cell lines. The interaction of Grb7 with the ErbB2 receptor is mediated via its Src homology 2 (SH2) domain. Whilst most SH2 domains exist as monomers, recently reported studies have suggested that the Grb7-SH2 domain exists as a homodimer. The self-association properties of the Grb7-SH2 domain were therefore studied using sedimentation equilibrium ultracentrifugation. Analysis of the data demonstrated that the Grb7-SH2 domain is dimeric with a dissociation constant of approximately 11 muM. We also demonstrate, using size-exclusion chromatography, that mutation of phenylalanine 511 to an arginine produces a monomeric form of the Grb7-SH2 domain. This mutation represents the first step in the engineering of a Grb7-SH2 domain with good solution properties for further biophysical and structural investigation.
Original languageEnglish
Pages (from-to)454 - 460
Number of pages7
JournalEuropean Biophysics Journal
Volume34
Issue number5
DOIs
Publication statusPublished - 2005

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