A new reporter cell line for studies with proteasome inhibitors in Trypanosoma brucei.
Mol Biochem Parasitol
; 227: 15-18, 2019 01.
Article
in En
| MEDLINE
| ID: mdl-30444978
ABSTRACT
A Trypanosoma brucei cell line is described that produces a visual readout of proteasome activity. The cell line contains an integrated transgene encoding an ubiquitin-green fluorescent protein (GFP) fusion polypeptide responsive to the addition of proteasome inhibitors. A modified version of T. brucei ubiquitin unable to be recognized by deubiquitinases (UbG76V) was fused to eGFP and constitutively expressed. The fusion protein is unstable but addition of the proteasome inhibitor lactacystin stabilizes it and leads to visually detectable GFP. This cell line can be widely used to monitor the efficiency of inhibitor treatment through detection of GFP accumulation in studies involving proteasome-mediated proteolysis, screening of proteasome inhibitors or other events related to the ubiquitin-proteasome pathway.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Trypanocidal Agents
/
Trypanosoma brucei brucei
/
Cell Line
/
Drug Evaluation, Preclinical
/
Proteasome Inhibitors
Type of study:
Evaluation_studies
Limits:
Humans
Language:
En
Journal:
Mol Biochem Parasitol
Year:
2019
Document type:
Article