Understanding Protein Mobility in Bacteria by Tracking Single Molecules

JOURNAL OF MOLECULAR BIOLOGY

卷:430

Review

Kapanidis, Achillefs N.¹  Uphoff, Stephan²  Stracy, Mathew² 

[1] Univ Oxford, Dept Phys, Clarendon Lab, Gene Machines Grp,Biol Phys Res Unit, Oxford OX1 3PU, England

[2] Univ Oxford, Dept Biochem, Oxford OX1 3QU, England

关键词: protein diffusion;    single-molecule tracking;    anomalous diffusion;    macromolecular crowding;    target search;   

DOI  :  10.1016/j.jmb.2018.05.002

来源: Elsevier

【 摘 要 】

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. (C) 2018 The Authors. Published by Elsevier Ltd.

【 授权许可】

Free