Work

Optimized protocol to detect protein UFMylation in cells and in vitro via immunoblotting

Year: 2022
Type: article
Abstract: Ubiquitin-fold modifier 1 (UFM1) system is a recently identified ubiquitin-like modification with essential biological functions. Similar to ubiquitination, the covalent conjugation of UFM1 (UFMylation) to target proteins involves a three-step enzymatic cascade catalyzed sequentially by UFM1-activating enzyme 5 (UBA5, E1), UFM1-conjugating enzyme 1 (UFC1, E2), and UFM1-specific ligase 1 (UFL1, E3). Here, we provide an optimized protocol adapted to previously reported methods for detecting the UFMylation of target protein in human cells and in vitro assays, respectively, with high reliability and reproducibility. For complete details on the use and execution of this protocol, please refer to Liu et al. (2020).
Language: English
Open Access status: gold
Estimated APC paid: $1,500
OpenAlex ID: W4205244831
FWCI: 0.898
Cites: 7
Cited by: 18
Related to: 10