Identification of newly translated thermo-sensitive proteins using pulse SILAC mass spectrometry and the GAL promoter system.
STAR Protoc
; 4(1): 102059, 2023 03 17.
Article
en En
| MEDLINE
| ID: mdl-36853680
ABSTRACT
Some newly translated proteins are more susceptible to misfolding and aggregation upon heat shock in comparison to other proteins. To study these newly translated thermo-sensitive proteins on a proteomic scale, we present here a protocol that combines pulse-SILAC with biochemical fractionation for mass spectrometry analysis, followed by an orthogonal validation protocol for selected candidates using the GAL promoter system in Saccharomyces cerevisiae. This approach can be further developed to study other stresses and specific post-translational modifications or adapted to mammalian cells. For complete details on the use and execution of this protocol, please refer to Zhu et al. (2022).1.
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Texto completo:
1
Base de datos:
MEDLINE
Asunto principal:
Proteómica
/
Fraccionamiento Químico
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
STAR Protoc
Año:
2023
Tipo del documento:
Article