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. 2015 Jul 30;290(38):23188–23200. doi: 10.1074/jbc.M115.648436

FIGURE 1.

FIGURE 1.

GluN2C C terminus interacts with various 14-3-3 isoforms, and binding is regulated by Ser-1096. A, schematic representation of yeast two-hybrid constructs used, including an alignment of GluN2A, GluN2B, and GluN2C. GluN2C Ser-1096 is indicated with an arrowhead and 14-3-3 binding motif is indicated with an arrow. The last 175 amino acids within the C terminus of GluN2A, GluN2B, or GluN2C were fused to LexA DNA binding domain. Each 14-3-3 isoform was fused to Gal4 activation domain and used for subsequent yeast two-hybrid binding assay. B, yeast were co-transformed with LexA-GluN2C WT C terminus and either Gal4 vector or Gal4-14-3-3Inline graphic, Gal4-14-3-3ζ, Gal4-14-3-3τ, Gal4- 14-3-3ϵ, Gal4-14-3-3η, Gal4-14-3-3β, or Gal4-14-3-3σ, and growth was evaluated on appropriate yeast selection medium. C, yeast were co-transformed with either LexA-GluN2C S1096A or LexA-GluN2C S1144A and either Gal4 vector or Gal4-14-3-3ϵ, τ, η, Inline graphic, ζ, or β, and growth was evaluated on appropriate yeast selection medium. B and C, results shown are 10-fold serial dilutions of yeast cells. All data shown are representative of at least three independent experiments.