LW
Lai Chu Wu
  • Rheumatology & Immunology, Ohio State University Wexner Medical Center, Columbus
In vitro Crosslinking Reactions and Substrate Incorporation Assays for The Identification of Transglutaminase-2 Protein Substrates
转谷氨酰胺酶-2蛋白底物的体外交联反应和底物掺入鉴定
作者:William L. Willis, Abigail Foster, Caitlin Henry, Lai Chu Wu and Wael Jarjour日期:06/20/2020,浏览量:2970,Q&A: 0
Transglutaminase (TG2) catalyzes protein crosslinking between glutamyl and lysyl residues. Catalytic activity occurs via a transamidation mechanism resulting in the formation of isopeptide bonds. Since TG2-mediated transamidation is of mechanistic importance for a number of biological processes, assays that enable rapid and efficient identification and characterization of candidate substrates are an important first-step to uncovering the function of crosslinked proteins. Herein we describe an optimized and flexible protocol for in vitro TG2 crosslink reactions and substrate incorporation assays. We have previously employed these techniques in the identification of the protein high mobility group box 1 (HMGB1) as a TG2 substrate. However, the protocol can be adapted for identification of any candidate transamidation substrate.