Quantifying Multistate Cytoplasmic Molecular Diffusion in Bacterial Cells via Inverse Transform of Confined Displacement Distribution
Year: 2015
Type: article
Abstract: Single-molecule tracking (SMT) of fluorescently tagged cytoplasmic proteins can provide valuable information on the underlying biological processes in living cells via subsequent analysis of the displacement distributions; however, the confinement effect originated from the small size of a bacterial cell skews the protein's displacement distribution and complicates the quantification of the intrinsic diffusive behaviors. Using the inverse transformation method, we convert the skewed displacement distribution (for both 2D and 3D imaging conditions) back to that in free space for systems containing one or multiple (non)interconverting Brownian diffusion states, from which we can reliably extract the number of diffusion states as well as their intrinsic diffusion coefficients and respective fractional populations. We further demonstrate a successful application to experimental SMT data of a transcription factor in living E. coli cells. This work allows a direct quantitative connection between cytoplasmic SMT data with diffusion theory for analyzing molecular diffusive behavior in live bacteria.
Authors: Tai-Yen Chen, Won Jung, Ace George Santiago, Feng Yang, Łukasz Krzemiński, Peng Chen
Institution: Cornell University
Language: English
Subfield: Biophysics
Domain: Life Sciences
Open Access status: hybrid
Awards: Single Molecule Optically Resonant NanoTweezers for the study of Intracellular Me, G3766716424, G3768959624, G7247574548, Single-molecule Imaging of Tripartite Membrane Efflux Pumps in Live Bacteria, Unusual mechanisms of metal regulation in bacteria: from single molecules to single cells to cell communities, G947352956
OpenAlex ID: W2432296310