A Chemical and Enzymatic Approach to Study Site-Specific Sumoylation

• DOI: 10.1371/journal.pone.0143810
• Publication Date: 2015-12-03T18:46:48Z
• Authors: Claudio P. Albuqu...; Eyan Yeung; Shawn Ma; Ting Fu; Kevin D. Corbett; Huilin Zhou
• Keywords: SUMO enzymes; Proteome

A variety of cellular pathways are regulated by protein modifications with ubiquitin-family proteins. SUMO, the Small Ubiquitin-like MOdifier, is covalently attached to lysine on target proteins via a cascade reaction catalyzed E1, E2, and E3 enzymes. major barrier understanding diverse regulatory roles SUMO has been lack suitable methods identify sumoylation sites. Here we developed mass-spectrometry (MS) based approach combining chemical enzymatic We applied this method analyze auto-sumoylation E1 enzyme in vitro compared it GG-remnant using Smt3-I96R as substrate. further examined effect smt3-I96R mutation vivo performed proteome-wide analysis sites Saccharomyces cerevisiae. To validate these findings, confirmed several Aos1 Uba2 vivo. Together, results demonstrate that our for identifying provides useful tool combination allows detailed