Open AccessDevelopment of Binding Assays for the SH2 Domain of Grb7 and Grb2 Using Fluorescence Polarization
Author(s) -
Jean-Philippe Luzy,
Huixiong Chen,
Brunilde Gril,
WangQing Liu,
Michel Vidal,
Dominique Perdereau,
Anne-Françoise Burnol,
Christiane Garbay
Publication year - 2008
Publication title -
slas discovery
Language(s) - English
Resource type - Journals
eISSN - 2472-5560
pISSN - 2472-5552
DOI - 10.1177/1087057107312124
Subject(s) - sh2 domain , fluorescence anisotropy , grb2 , affinities , peptidomimetic , proto oncogene tyrosine protein kinase src , chemistry , signal transducing adaptor protein , biochemistry , tyrosine , biology , phosphorylation , peptide , membrane
Adaptor proteins Grb7 and Grb2 have been implicated as being 2 potential therapeutic targets in several human cancers, especially those that overexpress ErbB2. These 2 proteins contain both a SH2 domain (Src homology 2) that binds to phosphorylated tyrosine residues contained within ErbB2 and other specific protein targets. Two assays based on enzyme-linked immunosorbent assay and fluorescence polarization methods have been developed and validated to find and rank inhibitors for both proteins binding to the pY(1139). Fluorescence polarization assays allowed the authors to determine quickly and reproducibly affinities of peptides from low nanomolar to high micromolar range and to compare them directly for Grb7 and Grb2. As a result, the assays have identified a known peptidomimetic Grb2 SH2 inhibitor (mAZ-pTyr-(alphaMe)pTyr-Asn-NH(2)) that exhibits the most potent affinity for the Grb7 SH2 domain described to date.