A non-fluorescent HaloTag blocker for improved measurement and visualization of protein synthesis in living cells.

ABSTRACT: Background: HaloTag is a modified bacterial enzyme that binds rapidly and irreversibly to an array of synthetic ligands, including chemical dyes. When expressed in live cells in conjunction with a protein of interest, HaloTag can be used to study protein trafficking, synthesis, and degradation. For instance, sequential HaloTag labeling with spectrally separable dyes can be used to separate preexisting protein pools from proteins newly synthesized following experimental manipulations or the passage of time. Unfortunately, incomplete labeling by the first dye, or labeling by residual, trapped dye pools can confound interpretation. Methods: Labeling specificity of newly synthesized proteins could be improved by blocking residual binding sites. To that end, we synthesized a non-fluorescent, ce

SUBMITTER: Cohen LD 

PROVIDER: S-EPMC7255903 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature