Throughout the cell, motor proteins work together to drive numerous molecular processes and functions. For example, ensembles of myosin motors collectively transport vesicles and organelles, maintain membrane homeostasis, and drive muscle contraction. Studying these motors in groups has become increasingly important with work demonstrating the emergence of ensemble behavior distinct from individual motor behavior. One powerful technique that has been used in the last decade is DNA nanotechnology, which provides precise control over spacing and organization of patterned motor proteins. Until recently, however, most studies combining DNA nanostructures and molecular motors have been confined to discrete DNA structures with limited attachment points for motor proteins. In this chapter, we describe a new approach for making synthetic motor filaments using DNA nanotubes. We present methods for preparing myosin VI-labeled nanotubes and testing these nanotubes using a general in vitro motility setup. Overall, these nanotubes can easily be used to study other large ensembles of molecular motors, such as muscle myosin or ciliary dynein, both proteins that work in large motor ensembles to drive key cellular functions.
|Original language||English (US)|
|Title of host publication||Methods in Molecular Biology|
|Publisher||Humana Press Inc.|
|Number of pages||9|
|State||Published - 2018|
|Name||Methods in Molecular Biology|
Bibliographical noteFunding Information:
This work was supported by the American Heart Association Scientist Development Grant (13SDG14270009) and the NIH (1DP2 CA186752-01 and 1-R01-GM-105646-01-A1) to SS. RFS is a Life Sciences Research Foundation postdoctoral fellow.
- DNA nanotechnology
- In vitro motility
- Molecular motors
- Myosin VI