Harnessing Split-Inteins as a Tool for the Selective Modification of Surface Receptors in Live Cells.

ABSTRACT: Biochemical studies of integral membrane proteins are often hampered by low purification yields and technical limitations such as aggregation causing in vitro manipulations to be challenging. The ability of controlling proteins in live cells bypasses these limitations while broadening the scope of accessible questions owing to the proteins being in their native environment. Here we take advantage of the intein biorthogonality to mammalian systems, site specificity, fast kinetics, and auto-processing nature as an attractive option for modifying surface proteins. Using EGFR as a model, we demonstrate that the split-intein pair Ava^N /Npu^C can be used to efficiently and specifically modify target membrane proteins with a synthetic adduct for downstream live cell application.

SUBMITTER: Ray DM 

PROVIDER: S-EPMC9977608 | biostudies-literature | 2023 Feb

REPOSITORIES: biostudies-literature