On-Chip SUMOylation for high-throughput proteome analysis

Abstract

This paper describes the novel strategy for on chip SUMOylation with using Spreeta sensor chip. It is clear that SUMOylation, one of the post translational modification, is very important cellular event in due consideration that SUMOylation regulates diverse cellular functions. Although, a small numbers of SUMO target proteins identified recently, it is obvious that the speed of discovering SUMO substrates will be accelerated when the high-throughput proteomic approaches apply to target screening. My study is based on that the Ubc9 is an essential SUMO conjugation enzyme to bind proteins including SUMO substrates. The protein chip immobilized with Ubc9 recognizes primarily Ubc9 binding proteins out of protein poll. Then, SUMOylation phenomenon as a specific sensorgram pattern was observed when GST tagged RanGAP1 and p53 proteins were applied. And then, the SUMO modified proteins were collected for qualitative analysis with mass spectroscopy. Also, the specific sensorgram pattern was not detected when GST tagged Ref-1 was layered on under same condition. It is believed that the data performed under in vivo is more reliable than the one under in vitro, because the SUMO modification is dynamic, with substrates undergoing rapid conjugation and deconjugation. But, it is significant work actualizing biological process engaging several protein complexes with such exemplary model system as SUMOylation on the Spreeta sensor chip and proposes new strategy to identify SUMO substartes by biomolecular interaction analysis mass spectrometry(BIA/MS).