SUMO (small ubiquitin-like modifier) chip for analysis of SUMO-conjugation to a target protein

Abstract

The analysis of protein interactions and modifications in a microarray format has become the focus of great interest in researchers in drug discovery, diagnostics, and cell biology, due to the advent of rapid and high-throughput biological assays requiring only a small amount of sample. Herein, we describe a chip-based analysis of sumoylation, the post-translational modification (PTM) process involving the covalent attachment of the small ubiquitin-like modifier (SUMO) family protein to a target protein in a mammalian cell. The protein was expressed using an in vitro translation system and immobilized directly onto a glass slide using epitope tags fused to the protein. In this paper, the microarray-based analysis technique was shown to be a useful strategy for the identification of SUMO target proteins from preexisting protein pools and proteome arrays, in a high-throughput manner.