Solid-phase binding assays of peptides using EGFP-Src SH2 domain fusion protein and biotinylated Src SH2 domain

Guofeng Ye, Marina Ayrapetov, Nguyen Hai Nam, Gongqin Sun, Keykavous Parang

Research output: Contribution to journalArticlepeer-review

Abstract

Two solid-phase binding assays were designed and evaluated for their potential use in comparing the affinity of peptides to the Src SH2 domain. Resin beads attached to peptides were incubated with the enhanced green fluorescence protein(EGFP)-Src SH2 domain fusion protein or the biotinylated Src SH2 domain and extensively washed. The beads-attached tetrapeptides with high affinities to the EGFP-Src SH2 domain showed more fluorescence intensity than those beads containing tetrapeptides with weak binding affinities, as shown by fluorescence microscopy and fluorescence imaging system. Only the beads attached to pYEEI produced a dark purple color on incubation of the beads, respectively, with the biotinylated Src kinases SH2 domain, alkaline phosphatase-coupled streptavidin, and BCIP/NBT. These solid-phase binding assays may have potential applications for the screening of peptides for the Src kinases SH2 domains.

Original languageEnglish (US)
Pages (from-to)4994-4997
Number of pages4
JournalBioorganic and Medicinal Chemistry Letters
Volume15
Issue number22
DOIs
StatePublished - Nov 15 2005
Externally publishedYes

Keywords

  • Binding assays
  • Biotinylation
  • Fluorescence
  • Solid-phase
  • Src SH2 domain

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Organic Chemistry
  • Drug Discovery
  • Pharmaceutical Science

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