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2050-01-01
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A NEW REPORTER CELL LINE FOR STUDIES WITH PROTEASOME INHIBITORS IN TRYPANOSOMA BRUCEI
Avaliação Pré-Clínica de Medicamentos / métodos
Genes Reporter
Proteínas de Fluorescência Verde / genética
Proteínas de Fluorescência Verde / metabolismo
Humanos
Complexo de Endopeptidases do Proteassoma / genética
Complexo de Endopeptidases do Proteassoma / metabolismo
Inibidores de Proteassoma / farmacologia
Tripanossomicidas / farmacologia
Trypanosoma brucei brucei / efeitos dos fármacos
Trypanosoma brucei brucei / enzimologia
Trypanosoma brucei brucei / genética
Tripanossomíase Africana / parasitologia
Ubiquitina / metabolismo
Affilliation
Fundação Oswaldo Cruz. Instituto Aggeu Magalhães. Recife, PE, Brasil / University of Cambridge. Department of Biochemistry. Cambridge, United Kingdom.
Fundação Oswaldo Cruz. Instituto Aggeu Magalhães. Recife, PE, Brasil.
University of Cambridge. Department of Biochemistry. Cambridge, United Kingdom.
Fundação Oswaldo Cruz. Instituto Aggeu Magalhães. Recife, PE, Brasil.
University of Cambridge. Department of Biochemistry. Cambridge, United Kingdom.
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.
DeCS
Linhagem CelularAvaliação Pré-Clínica de Medicamentos / métodos
Genes Reporter
Proteínas de Fluorescência Verde / genética
Proteínas de Fluorescência Verde / metabolismo
Humanos
Complexo de Endopeptidases do Proteassoma / genética
Complexo de Endopeptidases do Proteassoma / metabolismo
Inibidores de Proteassoma / farmacologia
Tripanossomicidas / farmacologia
Trypanosoma brucei brucei / efeitos dos fármacos
Trypanosoma brucei brucei / enzimologia
Trypanosoma brucei brucei / genética
Tripanossomíase Africana / parasitologia
Ubiquitina / metabolismo
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