Deciphering a hexameric protein complex with Angstrom optical resolution.

ABSTRACT: Cryogenic optical localization in three dimensions (COLD) was recently shown to resolve up to four binding sites on a single protein. However, because COLD relies on intensity fluctuations that result from the blinking behavior of fluorophores, it is limited to cases where individual emitters show different brightness. This significantly lowers the measurement yield. To extend the number of resolved sites as well as the measurement yield, we employ partial labeling and combine it with polarization encoding in order to identify single fluorophores during their stochastic blinking. We then use a particle classification scheme to identify and resolve heterogenous subsets and combine them to reconstruct the three-dimensional arrangement of large molecular complexes. We showcase this method (po

SUBMITTER: Mazal H 

PROVIDER: S-EPMC9142145 | biostudies-literature | 2022 May

REPOSITORIES: biostudies-literature