Development of luciferase-based highly sensitive reporters that detect ER-associated protein biogenesis abnormalities.

ABSTRACT: Localization to the endoplasmic reticulum (ER) and subsequent disulfide bond formation are crucial processes governing the biogenesis of secretory pathway proteins in eukaryotes. Hence, comprehending the mechanisms underlying these processes is important. Here, we have engineered firefly luciferase (FLuc) as a tool to detect deficiencies in these processes within mammalian cells. To achieve this, we introduced multiple cysteine substitutions into FLuc and targeted it to the ER. The reporter exhibited FLuc activity in response to defects in protein localization or disulfide bond formation within the ER. Notably, this system exhibited outstanding sensitivity, reproducibility, and convenience in detecting abnormalities in these processes. We applied this system to observe a protein translocat

SUBMITTER: Kadokura H 

PROVIDER: S-EPMC11564982 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature