Titlebook: Dynein; Methods and Protocol Steven M. Markus Book 2023 The Editor(s) (if applicable) and The Author(s), under exclusive license to Springe

种类

ungainly

https://doi.org/10.1007/978-3-642-57192-3es. We provide updated and optimized protocols for the expression and purification of a tail-truncated single-headed . dynein, for polarity-marked MT polymerization, and for the non-covalent attachment of MTs to cover glass surfaces for the measurement of dynein-MT detachment forces.

fatty-streak

Employing Live-Cell Imaging to Study Motor-Mediated Transport users to examine the net effect of motors and motor-regulatory proteins on organelle distribution. Together, these methods open the door to motility experiments that more fully interrogate the complex cellular environment.

百灵鸟

Measurements of the Force-Dependent Detachment Rates of Cytoplasmic Dynein from Microtubuleses. We provide updated and optimized protocols for the expression and purification of a tail-truncated single-headed . dynein, for polarity-marked MT polymerization, and for the non-covalent attachment of MTs to cover glass surfaces for the measurement of dynein-MT detachment forces.

宠爱

改正

Using Optogenetics to Spatially Control Cortical Dynein Activity in Mitotic Human Cellstion and manipulate downstream events. In this chapter, I will present methods for locally activating cytoplasmic dynein at the mitotic cell cortex in human cells, with a focus on how to generate knock-in cell lines and set up a microscope system.

GRE

Reconstitution of Dynein/Dynactin Transport Using Recombinant Dynein adaptors, and endogenous dynactin and detail a protocol for the single-molecule motility assay. In this assay, microtubules are first immobilized on a coverslip. A fluorescently labeled dynein/dynactin/cargo adaptor complex is then added, allowing for the measurement of key motility parameters as the complex walks along the microtubule.

hermetic

Fast and Easy Transient Mammalian Cell Expression and Purification of Cytoplasmic Dyneinpurification for high-throughput protein engineering. Mammalian cell expression might be generally considered to be a laborious process, but with recent technology and some simple inexpensive custom-built labware, dynein expression and purification from mammalian cells can be fast and easy.

achlorhydria

https://doi.org/10.1007/978-3-642-57192-3external factors affect the microtubule-detachment rate and velocity of dynein. We also briefly discuss the remaining challenges and future directions of optical trapping studies of dyneins and other microtubule-based motors.

胡言乱语

Studying Dynein Mechanochemistry with an Optical Trapexternal factors affect the microtubule-detachment rate and velocity of dynein. We also briefly discuss the remaining challenges and future directions of optical trapping studies of dyneins and other microtubule-based motors.