High-Resolution Tracking of Dynein-Dynactin-BicD2 Complexes

Qingzhou Feng, Allison M. Gicking, William O. Hancock

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

The adapter dynactin and the activator BicD2 associate with dynein to form the highly motile dynein-dynactin-BicD2 (DDB) complex. In single-molecule assays, DDB displays processive runs, diffusive episodes, and transient pauses. The switching rates and durations of the different phases can be determined by tracking gold nanoparticle-labeled DDB complexes with interferometric scattering (iSCAT) microscopy and using an algorithm to separate out different motility phases. Here we describe methods for purifying DDB complexes from brain lysate, labeling with gold nanoparticles, imaging by iSCAT, and analyzing the resulting trajectories.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages177-186
Number of pages10
DOIs
StatePublished - 2023

Publication series

NameMethods in Molecular Biology
Volume2623
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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