Affordable Access

deepdyve-link
Publisher Website

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

Authors
  • Zhou, Junzhi1
  • Liang, Qian1
  • Dong, Maogong1
  • Ma, Xiaohe1
  • Jin, Yaqi1
  • Guan, Di1
  • Liu, Jiang1
  • Wang, Miao1
  • Cong, Yu-Sheng1
  • 1 Key Laboratory of Aging and Cancer Biology of Zhejiang Province, School of Basic Medical Sciences, Hangzhou Normal University, Hangzhou, China. , (China)
Type
Published Article
Journal
STAR Protocols
Publisher
Elsevier
Publication Date
Mar 18, 2022
Volume
3
Issue
1
Pages
101074–101074
Identifiers
DOI: 10.1016/j.xpro.2021.101074
PMID: 35036955
Source
Medline
Keywords
Language
English
License
Unknown

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). © 2021 The Authors.

Report this publication

Statistics

Seen <100 times