FIGURE 2.

Light chain binding to NMDA receptor subunits does not require added magnesium. A direct comparison of light chain binding to the carboxyl terminus of NR1 and the neck region myosin IIB heavy chain revealed that NR1-RLC interactions are less sensitive to the effects of Mg²⁺. The carboxyl tail of NR1-(834-938) and the neck region of non-muscle myosin heavy chain IIB-(771-843), each fused to GST, were immobilized on glutathione-Sepharose beads for use in GST pulldown experiments. GST fusion proteins or GST alone were used as affinity matrices to examine interactions with purified, recombinant myosin RLC in the absence (lanes 2-4) or presence (lanes 5-7) of 1 mm Mg²⁺. Assays were initiated by the addition of 300 nm myosin RLC and incubated with rotation for 2 h at 4 °C. Immune complexes were revealed using an anti-T-7 antibody (Novagen) to detect recombinant myosin RLC. Figure is representative of three independent experiments.