Identification of striated muscle activator of Rho signaling (STARS) as a novel calmodulin target by a newly developed genome-wide screen

Identification and Biochemical Characterization of High Mobility Group Protein 20A as a Novel Ca2+/S100A6 Target

“…Protein array plates (Protein Active Array ® , 1536-well format × 27 plates) containing 19,676 recombinant human proteins in duplicate, generated by wheat germ cell-free protein synthesis, were constructed by CellFree Sciences [17], and the screening for protein-protein interactions was performed by incubation with 0.36 µg/mL biotinylated S100A6, followed by a streptavidin-horseradish peroxidase-mediated enhanced chemiluminescence (ECL) detection of S100A6 binding with the ChemiDoc™ XRS (Bio-Rad Laboratories, Hercules, CA, USA), as previously reported [18].…”

“…To comprehensively analyze target proteins of EF-hand Ca 2+ -binding proteins, including calmodulin and S100 proteins, we developed a novel genome-wide screening method based on the protein-protein interactions using the Protein Active Array ® (CellFree Sciences, Ehime, Japan), carrying 19,676 recombinant glutathione s-transferase (GST)-fusion proteins [17]. The use of this method resulted in the identification of the striated muscle activator of Rho signaling as a calmodulin interactant, and FOR-20 (FOP-related protein of 20 kDa) as a novel S100A6-interacting protein [18].…”

Isogenic human pluripotent stem cell disease models reveal ABRA deficiency underlies cTnT mutation-induced familial dilated cardiomyopathy

“…2 H and 2 I). Previous study showed that the Ca 2+ -dependent protein calmodulin directly binds to the N-terminus of ABRA and negatively regulates muscle gene expression induced by the ABRA-serum response factor (SRF) pathway (Furuya et al, 2016 ), thereby linking Ca 2+ -signaling to ABRA-mediated gene expression. Immunoprecipitation assays showed that calmodulin binds to ABRA in cardiomyocytes (Fig.…”