Thioether editing generally increases the photostability of rhodamine dyes on self-labeling tags.

ABSTRACT: Self-labeling protein tags are widely used in advanced bioimaging where dyes with high-photon budgets outperform their fluorescent protein counterparts. Further increasing the emitted photon numbers of dye-tag systems is actively pursued by both new fluorophore chemistry and protein engineering. By scrutinizing the protein microenvironment of fluorophores, here we propose that proximal thioether groups negatively affect the photostability of the dye-tag system. We attribute the disparity in photostability of rhodamine dyes on HaloTag, SNAP-tag, and TMP-tag3 to the influence of the inherent thioether linkage within the SNAP-tag and TMP-tag3. This photochemical pathway leads us to further devise tags with higher photostability. We first show that rhodamine dyes on TMP-tag3.1, which employs a

SUBMITTER: Ling J 

PROVIDER: S-EPMC12318191 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature