title: Reconstitution of microtubule-based motility using cell extracts.
abstract: Long-range intracellular transport of organelles driven by kinesin and dynein motor proteins depends on additional cellular factors including adaptors and scaffolding proteins. While single-molecule studies of the motility of purified motor proteins have been a powerful approach, these assays are not fully representative of the complex interactions that occur in a cellular environment. To gain insights into the functioning of adaptor proteins that work in concert with motors proteins, motility assays in cell extracts have been developed. These assays are an attractive means to begin to dissect the roles of additional factors in motor-driven transport. Further, this system can be easily manipulated to study this process in different physiological environments. Here we describe in vitro reconstitution of motor-driven motility along microtubules in cell extracts, followed by considerations for data analysis and how these assays can be powerful in informing our understanding of basic cellular processes.
authors.affiliation: Department of Physiology and the Pennsylvania Muscle Institute, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, USA; Department of Biology Graduate Group, School of Arts and Sciences at the University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Swathi
authors.initials: S
authors.lastname: Ayloo
authors.affiliation: Department of Physiology and the Pennsylvania Muscle Institute, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Erika L F
authors.initials: EL
authors.lastname: Holzbaur
mesh_terms: D000818:Animals; D001692:Biological Transport / Q000502:physiology*; D019556:COS Cells; D002457:Cell Extracts; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology*; D002522:Chlorocebus aethiops; D004398:Dyneins / Q000378:metabolism*; D016547:Kinesins / Q000378:metabolism*; D007734:Kymography / Q000379:methods; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism*; D015388:Organelles / Q000378:metabolism
chemical_list: D002457:Cell Extracts; D008869:Microtubule-Associated Proteins; D004398:Dyneins; D016547:Kinesins
keywords: Cell extract; Dynein; Kinesin; Microtubule; Motility; Motor proteins