Detection and Identification of Protein Interactions of S100 Proteins by ProteinChip Technology

Abstract

The aim of this work was to establish an approach for identification of protein interactions. This assay used an anti-S100A8 antibody coupled on beads and incubated with cell extract. The bead eluates were analyzed using ProteinChip technology and subsequently subjected to an appropriate digestion. Molecular masses of digestion fragments were determined by SELDI−MS, and database analysis revealed S100A10 as interacting protein. This result was confirmed by co-immunoprecipitation and immunocapturing. Using S100A10 as new bait, a specific interaction with S100A7 was detectable. Keywords: S100 proteins • protein−protein interaction assay • surface-enhanced laser desorption/ionization−mass spectrometry (SELDI−MS) • immunoprecipitation