Sulfo-SMCC Prevents Annealing of Taxol-Stabilized Microtubules In Vitro.

Published

Journal Article

Microtubule structure and functions have been widely studied in vitro and in cells. Research has shown that cysteines on tubulin play a crucial role in the polymerization of microtubules. Here, we show that blocking sulfhydryl groups of cysteines in taxol-stabilized polymerized microtubules with a commonly used chemical crosslinker prevents temporal end-to-end annealing of microtubules in vitro. This can dramatically affect the length distribution of the microtubules. The crosslinker sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate, sulfo-SMCC, consists of a maleimide and an N-hydroxysuccinimide ester group to bind to sulfhydryl groups and primary amines, respectively. Interestingly, addition of a maleimide dye alone does not show the same interference with annealing in stabilized microtubules. This study shows that the sulfhydryl groups of cysteines of tubulin that are vital for the polymerization are also important for the subsequent annealing of microtubules.

Full Text

Duke Authors

Cited Authors

  • Prabhune, M; von Roden, K; Rehfeldt, F; Schmidt, CF

Published Date

  • January 2016

Published In

Volume / Issue

  • 11 / 8

Start / End Page

  • e0161623 -

PubMed ID

  • 27561096

Pubmed Central ID

  • 27561096

Electronic International Standard Serial Number (EISSN)

  • 1932-6203

International Standard Serial Number (ISSN)

  • 1932-6203

Digital Object Identifier (DOI)

  • 10.1371/journal.pone.0161623

Language

  • eng