S100A6与靶蛋白在体外和活细胞中的相互作用。

Interaction of S100A6 with Target Proteins In Vitro and in Living Cells.

作者信息

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

机构信息

Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, Kita-ku, Okayama, Japan.

Ehime Prefectural University of Health Sciences, Iyo-gun, Ehime, Japan.

出版信息

Methods Mol Biol. 2019;1929:367-377. doi: 10.1007/978-1-4939-9030-6_23.

DOI:10.1007/978-1-4939-9030-6_23

PMID:30710285

Abstract

S100A6 is a member of the EF-hand Ca-binding protein family, which plays important roles in a wide variety of Ca 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 signaling. Moreover, they may be used to evaluate other physical S100/target interactions.

摘要

S100A6是EF手型钙结合蛋白家族的成员，在细胞内多种钙信号传导以及病理生理状况中发挥重要作用。在此，我们描述了用于体外评估S100A6与多种靶蛋白相互作用的分析方案，包括生物素化S100A6覆盖法、谷胱甘肽-S-转移酶（GST）沉淀法、表面等离子体共振法以及活细胞中的GST沉淀分析。这些方法将阐明S100A6/靶标相互作用的详细分子机制，并进一步增进我们对S100A6介导的钙信号传导生理意义的理解。此外，它们可用于评估其他S100/靶标之间的物理相互作用。