In vitro FRET- and fluorescence-based assays to study protein conformation and protein-protein interactions in mitosis

Stephanie C. Ems-McClung, Claire E. Walczak

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Proper cell division and the equal segregation of genetic material are essential for life. Cell division is mediated by the mitotic spindle, which is composed of microtubules (MTs) and MT-associated proteins that help align and segregate the chromosomes. The localization and characterization of many spindle proteins have been greatly aided by using GFP-tagged proteins in vivo, but these tools typically do not allow for understanding how their activity is regulated biochemically. With the recent explosion of the pallet of GFP-derived fluorescent proteins, fluorescence-based biosensors are becoming useful tools for the quantitative analysis of protein activity and protein-protein interactions. Here, we describe solution-based Förster resonance energy transfer (FRET) and fluorescence assays that can be used to quantify protein-protein interactions and to characterize protein conformations of MT-associated proteins involved in mitosis.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages93-122
Number of pages30
DOIs
StatePublished - Jan 1 2020
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume2101
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Fluorescence
  • FRET
  • Kinesin
  • Microtubule affinity
  • Mitosis
  • Protein conformation
  • Protein-protein interactions

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Fingerprint Dive into the research topics of 'In vitro FRET- and fluorescence-based assays to study protein conformation and protein-protein interactions in mitosis'. Together they form a unique fingerprint.

  • Cite this

    Ems-McClung, S. C., & Walczak, C. E. (2020). In vitro FRET- and fluorescence-based assays to study protein conformation and protein-protein interactions in mitosis. In Methods in Molecular Biology (pp. 93-122). (Methods in Molecular Biology; Vol. 2101). Humana Press Inc.. https://doi.org/10.1007/978-1-0716-0219-5_7