Open AccessThe fluorescent protein stability assay: an efficient method for monitoring intracellular protein stability
Author(s) -
Armelle Roisin,
Samuel Buchsbaum,
Vincent Mocquet,
Pierre Jalinot
Publication year - 2021
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2021-0032
Subject(s) - green fluorescent protein , intracellular , protein stability , translation (biology) , microbiology and biotechnology , protein biosynthesis , fluorescence , stability (learning theory) , protein expression , protein subcellular localization prediction , biology , chemistry , biophysics , computational biology , biochemistry , messenger rna , computer science , gene , physics , quantum mechanics , machine learning
The stability of intracellular proteins is highly variable, from a few minutes to several hours, and can be tightly regulated to respond to external and internal cellular environment changes. Several techniques can be used to study the stability of a specific protein, including pulse-chase labeling and blocking of translation. Another approach that has gained interest in recent years is fusing a protein of interest to a fluorescent reporter. In this report, the authors present a new version of this approach aimed at optimizing expression and comparison of the two reporter proteins. The authors show that the system works efficiently in various cells and can be useful for studying changes in protein stability and assessing the effects of drugs.