title: Development of binding assays for the SH2 domain of Grb7 and Grb2 using fluorescence polarization.
abstract: 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.
authors.affiliation: INSERM U648, Laboratoire de Pharmacochimie Moléculaire et Cellulaire, UFR Biomédicale, Université Paris Descartes, Paris, France.
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Luzy
authors.forename: Huixiong
authors.initials: H
authors.lastname: Chen
authors.forename: Brunilde
authors.initials: B
authors.lastname: Gril
authors.forename: Wang-Qing
authors.initials: WQ
authors.lastname: Liu
authors.forename: Michel
authors.initials: M
authors.lastname: Vidal
authors.forename: Dominique
authors.initials: D
authors.lastname: Perdereau
authors.forename: Anne-Françoise
authors.initials: AF
authors.lastname: Burnol
authors.forename: Christiane
authors.initials: C
authors.lastname: Garbay
mesh_terms: D001667:Binding, Competitive; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D016231:Fluorescence Polarization Immunoassay / Q000379:methods; D051380:GRB2 Adaptor Protein / Q000378:metabolism*; D051382:GRB7 Adaptor Protein / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D012680:Sensitivity and Specificity; D013379:Substrate Specificity; D018909:src Homology Domains
chemical_list: D051380:GRB2 Adaptor Protein; D051382:GRB7 Adaptor Protein; D004121:Dimethyl Sulfoxide