Measuring force generation within reconstituted microtubule bundle assemblies using optical tweezers

Autor: Omayma Al Azzam, Dana N. Reinemann, Cameron Lee Trussell
Rok vydání: 2021
Předmět:
Zdroj: Cytoskeleton. 78:111-125
ISSN: 1949-3592
1949-3584
DOI: 10.1002/cm.21678
Popis: Kinesins and microtubule associated proteins are critical to sustain life, facilitating cargo transport, cell division, and motility. To interrogate the mechanistic underpinnings of their function, these microtubule-based motors and proteins have been studied extensively at the single molecule level. However, a long-standing issue in the single molecule biophysics field has been how to investigate motors and associated proteins within a physiologically relevant environment in vitro. While the one motor/one filament orientation of a traditional optical trapping assay has revolutionized our knowledge of motor protein mechanics, this reductionist geometry does not reflect the structural hierarchy in which many motors work within the cellular environment. Here, we review approaches that combine the precision of optical tweezers with reconstituted ensemble systems of microtubules, microtubule associated proteins, and kinesins to understand how each of these unique elements work together to perform large scale cellular tasks, such as but not limited to building the mitotic spindle. Not only did these studies develop novel techniques for investigating motor proteins in vitro, but they also illuminate ensemble filament and motor synergy that helps bridge the mechanistic knowledge gap between previous single molecule and cell level studies. This article is protected by copyright. All rights reserved.
Databáze: OpenAIRE
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