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

Abstract

S100A6 is a member of the EF-hand Ca²⁺-binding protein family, which plays important roles in a wide variety of Ca²⁺ 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 Ca²⁺ signaling. Moreover, they may be used to evaluate other physical S100/target interactions.

Keywords: Ca2+-binding; EF-hand; FOR20; GST-precipitation assay; Protein phosphatase 5; Protein–protein interaction; S100A6; S100A6-overlay; SPR analysis; Transfection.