Use of the LC3B-fusion technique for biochemical and structural studies of proteins involved in the N-degron pathway.

ABSTRACT: The N-degron pathway, formerly the N-end rule pathway, is a protein degradation process that determines the half-life of proteins based on their N-terminal residues. In contrast to the well-established in vivo studies over decades, in vitro studies of this pathway, including biochemical characterization and high-resolution structures, are relatively limited. In this study, we have developed a unique fusion technique using microtubule-associated protein 1A/1B light chain 3B, a key marker protein of autophagy, to tag the N terminus of the proteins involved in the N-degron pathway, which enables high yield of homogeneous target proteins with variable N-terminal residues for diverse biochemical studies including enzymatic and binding assays and substrate identification. Intriguin

SUBMITTER: Kim L 

PROVIDER: S-EPMC7049975 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature