ArticleBiophysical journal2023
Dimerization of iLID optogenetic proteins observed using 3D single-molecule tracking in live E. coli.
Article in Biophysical journal, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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Who cites it
4 citing papers in PubMed, 6 citations in OpenAlex.
- Single-cell analysis and control of microbial systems using optogenetics.Current opinion in microbiology · 2026Review
- Synthetic Biomolecular Condensates: Design Principles and Applications.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Quantitative evaluation of methods to analyze motion changes in single-particle experiments.Nature communications · 2025Article
- Single-molecule tracking to determine the abundances and stoichiometries of freely-diffusing protein complexes in living cells: Past applications and future prospects.The Journal of chemical physics · 2023Review
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
3D single-molecule tracking microscopy has enabled measurements of protein diffusion in living cells, offering information about protein dynamics and cellular environments. For example, different diffusive states can be resolved and assigned to protein complexes of different size and composition. However, substantial statistical power and biological validation, often through genetic deletion of binding partners, are required to support diffusive state assignments. When investigating cellular processes, real-time perturbations to protein spatial distributions is preferable to permanent genetic deletion of an essential protein. For example, optogenetic dimerization systems can be used to manipulate protein spatial distributions that could offer a means to deplete specific diffusive states observed in single-molecule tracking experiments. Here, we evaluate the performance of the iLID optogenetic system in living E. coli cells using diffraction-limited microscopy and 3D single-molecule tracking. We observed a robust optogenetic response in protein spatial distributions after 488 nm laser activation. Surprisingly, 3D single-molecule tracking results indicate activation of the optogenetic response when illuminating with high-intensity light with wavelengths at which there is minimal photon absorbance by the LOV2 domain. The preactivation can be minimized through the use of iLID system mutants, and titration of protein expression levels.
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Registered trials
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