Fluorescent Probes for Monitoring Serine Ubiquitination.
Biochemistry
; 59(13): 1309-1313, 2020 04 07.
Article
em En
| MEDLINE
| ID: mdl-32207972
ABSTRACT
In a radical departure from the classical E1-E2-E3 three-enzyme mediated ubiquitination of eukaryotes, the recently described bacterial enzymes of the SidE family of Legionella pneumophila effectors utilize NAD+ to ligate ubiquitin onto target substrate proteins. This outcome is achieved via a two-step mechanism involving (1) ADP ribosylation of ubiquitin followed by (2) phosphotransfer to a target serine residue. Here, using fluorescent NAD+ analogues as well as synthetic substrate mimics, we have developed continuous assays enabling real-time monitoring of both steps of this mechanism. These assays are amenable to biochemical studies and high-throughput screening of inhibitors of these effectors, and the discovery and characterization of putative enzymes similar to members of the SidE family in other organisms. We also show their utility in studying enzymes that can reverse and inhibit this post-translational modification.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Serina
/
Proteínas de Bactérias
/
Bioquímica
/
Legionella pneumophila
/
Corantes Fluorescentes
Tipo de estudo:
Evaluation_studies
Idioma:
En
Revista:
Biochemistry
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
Estados Unidos