Interaction of S100A6 with target proteins in vitro and in living cells

Kyohei Sakane, Fuminori Yamaguchi, Mitsumasa Tsuchiya, Rina Kondo, Naoki Kanayama, Masaki Magari, Naoya Hatano, Ryoji Kobayashi, Hiroshi Tokumitsu

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

S100A6 is a member of the EF-hand Ca 2+ -binding protein family, which plays important roles in a wide variety of Ca 2+ signaling in the cells, as well as in pathophysiological conditions. Herein, we describe analytical protocols for evaluating the interaction of S100A6 with multiple target proteins in vitro, including biotinylated S100A6 overlay, glutathione-S-transferase (GST)-precipitation, surface plasmon resonance, and a GST-precipitation assay in living cells. These methods will elucidate the detailed molecular mechanisms of S100A6/target interactions and further improve our understanding of the physiological significance of S100A6-mediated Ca 2+ signaling. Moreover, they may be used to evaluate other physical S100/target interactions.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages367-377
Number of pages11
DOIs
Publication statusPublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1929
ISSN (Print)1064-3745

Keywords

  • Ca -binding
  • EF-hand
  • FOR20
  • GST-precipitation assay
  • Protein phosphatase 5
  • Protein–protein interaction
  • S100A6
  • S100A6-overlay
  • SPR analysis
  • Transfection

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Sakane, K., Yamaguchi, F., Tsuchiya, M., Kondo, R., Kanayama, N., Magari, M., Hatano, N., Kobayashi, R., & Tokumitsu, H. (2019). Interaction of S100A6 with target proteins in vitro and in living cells. In Methods in Molecular Biology (pp. 367-377). (Methods in Molecular Biology; Vol. 1929). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9030-6_23