Recording single motor proteins in the cytoplasm of mammalian cells.

Methods in Enzymology
Dawen CaiEdgar Meyhofer

Abstract

Biomolecular motors are central to the function and regulation of all cellular transport systems. The molecular mechanisms by which motors generate force and motion along cytoskeletal filaments have been mostly studied in vitro using a variety of approaches, including several single-molecule techniques. While such studies have revealed significant insights into the chemomechanical transduction mechanisms of motors, important questions remain unanswered as to how motors work in cells. To understand how motor activity is regulated and how motors orchestrate the transport of specific cargoes to the proper subcellular domain requires analysis of motor function in vivo. Many transport processes in cells are believed to be powered by single or very few motor molecules, which makes it essential to track, in real time and with nanometer resolution, individual motors and their associated cargoes and tracks. Here we summarize, contrast, and compare recent methodological advances, many relying on advanced fluorescent labeling, genetic tagging, and imaging techniques, that lay the foundation for groundbreaking approaches and discoveries. In addition, to illustrate the impact and capabilities for these methods, we highlight novel biological...Continue Reading

Citations

Mar 26, 2016·Frontiers in Cellular Neuroscience·Artur PadzikEleanor T Coffey
Mar 13, 2016·The EMBO Journal·Richard J McKenneyMinhajuddin Sirajuddin
Jul 21, 2012·Cell Biochemistry and Biophysics·Augusto BrunoValeria Levi

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