Reconstituting dynamic microtubule polymerization regulation by TOG domain proteins

Jawdat Al-Bassam

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

Abstract

Microtubules (MTs) polymerize from soluble αβ-tubulin and undergo rapid dynamic transitions to depolymerization at their ends. Microtubule-associated regulator proteins modulate polymerization dynamics in vivo by altering microtubule plus end conformations or influencing αβ-tubulin incorporation rates. Biochemical reconstitution of dynamic MT polymerization can be visualized with total internal reflection fluorescence (TIRF) microscopy using purified MT regulators. This approach has provided extensive details on the regulation of microtubule dynamics. Here, I describe a general approach to reconstitute MT dynamic polymerization with TOG domain microtubule regulators from the XMAP215/Dis1 and CLASP families using TIRF microscopy. TIRF imaging strategies require nucleation of microtubule polymerization from surface-attached, stabilized MTs. The approaches described here can be used to study the mechanism of a wide variety of microtubule regulatory proteins.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages131-148
Number of pages18
Volume540
ISBN (Print)9780123979247
DOIs
StatePublished - 2014

Publication series

NameMethods in Enzymology
Volume540
ISSN (Print)00766879
ISSN (Electronic)15577988

Keywords

  • CLASP
  • Microtubule dynamics
  • TIRF microscopy
  • TOG domains
  • Tubulin dimer
  • XMAP215/Dis1

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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  • Cite this

    Al-Bassam, J. (2014). Reconstituting dynamic microtubule polymerization regulation by TOG domain proteins. In Methods in Enzymology (Vol. 540, pp. 131-148). (Methods in Enzymology; Vol. 540). Academic Press Inc.. https://doi.org/10.1016/B978-0-12-397924-7.00008-X