Optimized protocol to detect protein UFMylation in cells and in vitro via immunoblotting.
STAR Protoc
; 3(1): 101074, 2022 03 18.
Article
em En
| MEDLINE
| ID: mdl-35036955
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).
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Enzimas Ativadoras de Ubiquitina
/
Enzimas de Conjugação de Ubiquitina
Tipo de estudo:
Prognostic_studies
Limite:
Humans
Idioma:
En
Revista:
STAR Protoc
Ano de publicação:
2022
Tipo de documento:
Article
País de afiliação:
China