Building a cell from its component parts by Jennifer Ross, Wallace F. Marshall

By Jennifer Ross, Wallace F. Marshall

This new quantity of Methods in mobilephone Biology appears at construction a telephone from its part elements. Chapters conceal such subject matters as engineering motor scaffolds, synthetic cytoskeletons, interconnected droplet networks and synthetic cells; construction cytoskeletal structures and synthetic actin cortex on pillar arrays; reconstituting membrane fission, actin bundles at membranes, actin cortex in droplets, dynein-dynactin mediated shipment shipping, brain structures, protein-lipid machines that remodel membranes, protein interactions and signaling on synthetic lipid bilayers. With state of the art fabric, this entire assortment is meant to steer researchers for years to come.

  • Covers sections on version structures and sensible stories, imaging-based techniques and rising studies
  • Chapters are written via specialists within the field
  • Cutting-edge material

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In this chapter, we describe our simple filament-gliding assay and then add modifications to the assay including adding cross-linking proteins and more filaments. Finally, we describe a system that recapitulates cell-like microtubule organizations similar to those found in mitosis using only three protein components. 2. Simple filament-gliding assay with kinesin-1 and microtubules 1. , 2013). We systematically add more microtubules or other types of associated proteins to probe how the patterns change in increased complexity.

23 24 CHAPTER 2 Microtubules, MAPs, and motor patterns Abstract Cells have an amazing ability to self-organize and rearrange their interiors. Such morphology changes are essential to cell development, division, and motility. The core of a cell’s internal organization lies with the cytoskeleton made of both microtubule and actin filaments with their associated proteins and ATP-utilizing enzymes. Despite years of in vitro reconstitution experiments, we still do not fully understand how the cytoskeleton can self-organize.

Moreover, the continuous flow of oil in principle opens the possibility to introduce (oil-soluble) drugs to the system to, for example, trigger microtubule polymerization. Note that the microfluidic chip can also be designed to produce elongated droplet in a vertical position, such that the poles of the droplets can be imaged at higher resolution. 19 20 CHAPTER 1 Microtubule-based cell polarity in fission yeast ACKNOWLEDGMENTS We thank Lutz Langguth for advice on thin layer optics. Magdalena Preciado-Lopez, Vanda Sunderlı´kova´, Cristina Manatschal, and Michel Steinmetz for help with the purification of proteins and for discussions.

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