In vitro reconstitution of small GTPase regulation

Abstract

Small GTPases play essential roles in the organization of eukaryotic cells. In recent years, it has become clear that their intracellular functions result from intricate biochemical networks of the GTPase and their regulators that dynamically bind to a membrane surface. Due to the inherent complexities of their interactions, however, revealing the underlying mechanisms of action is often difficult to achieve from in vivo studies. This review summarizes in vitro reconstitution approaches developed to obtain a better mechanistic understanding of how small GTPase activities are regulated in space and time.

Small GTPase signaling hubs at the surface of cellular membranes in physiology and disease

“…We are now delighted to present 13 articles that highlight how integrated structural, biochemical, and biophysical approaches have allowed to shed light on the mechanisms of fundamental functions of major members of this superfamily. Notably, these reviews show how several experimental approaches that were rarely used a decade ago, are now taking center stage in moving the field forward: reconstitutions of small GTPase systems in artificial membranes [5][6][7][8][9], single particle cryo-electron microscopy [5][6][7][9][10][11][12], 3D electron tomography reconstructions [12], or optogenetics [13]. Of these, Loose and coauthors present a great overview of the power and inherent difficulties of in vitro reconstitutions using biomimetic membranes, and how they have revealed in a quantitative manner the inner workings of intricate small GTPase networks, which would have been unattainable in vivo [8].…”