Understanding Protein Mobility in Bacteria by Tracking Single Molecules.

Journal of Molecular Biology
Achillefs N KapanidisMathew Stracy

Abstract

Protein diffusion is crucial for understanding the formation of protein complexes in vivo and has been the subject of many fluorescence microscopy studies in cells; however, such microscopy efforts are often limited by low sensitivity and resolution. During the past decade, these limitations have been addressed by new super-resolution imaging methods, most of which rely on single-particle tracking and single-molecule detection; these methods are revolutionizing our understanding of molecular diffusion inside bacterial cells by directly visualizing the motion of proteins and the effects of the local and global environment on diffusion. Here we review key methods that made such experiments possible, with particular emphasis on versions of single-molecule tracking based on photo-activated fluorescent proteins. We also discuss studies that provide estimates of the time a diffusing protein takes to locate a target site, as well as studies that examined the stoichiometries of diffusing species, the effect of stable and weak interactions on diffusion, and the constraints of large macromolecular structures on the ability of proteins and their complexes to access the entire cytoplasm.

Citations

Jan 30, 2020·Methods and Applications in Fluorescence·Angel ManceboElias M Puchner
Dec 4, 2019·Nature Chemical Biology·Anders S HansenXavier Darzacq
Sep 17, 2020·Biophysics Reviews·Pallavi KumariAntonio Benedetto
Dec 21, 2019·Scientific Reports·Yao-Kuan WangChien-Jung Lo
Feb 26, 2019·Nature Reviews. Microbiology·Oliver J Pambos, Achillefs N Kapanidis
Nov 12, 2020·International Journal of Molecular Sciences·Ostiane D'AugustinJuan Pablo Radicella
Mar 4, 2021·Journal of the Royal Society, Interface·Yuichi Itto, Christian Beck
Apr 30, 2019·Biophysical Journal·Julian RochaAndreas Gahlmann
Apr 17, 2021·Essays in Biochemistry·Dominika T Gruszka
Jun 2, 2021·EcoSal Plus·Maxime LerouxRodrigo Reyes-Lamothe
May 1, 2020·Biochemistry·Susanna K Yaeger-WeissSilvia Cavagnero
Jul 10, 2020·ACS Synthetic Biology·Leonard SchärfenMichael Schlierf
May 7, 2019·The Journal of Physical Chemistry. B·Niklas O JunkerJörg Fitter

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Methods Mentioned

BETA
fluorescence microscopy
fluorescence recovery after photobleaching
FRET

Software Mentioned

MINFLUX

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