Autodisplay of N-methyl-D-aspartate(NMDA)receptor[1]ligand binding domains(LBDs)was done by inserting the coding sequence for each LBD[2]into a plasmid for maximized autotransporter-mediated expression(pMATE)between t...Autodisplay of N-methyl-D-aspartate(NMDA)receptor[1]ligand binding domains(LBDs)was done by inserting the coding sequence for each LBD[2]into a plasmid for maximized autotransporter-mediated expression(pMATE)between the coding sequences of signal peptide and translocator domain[3](Figs.1A and S1).Surface display and functionality of LBDs was confirmed by protease accessibility and radioligand binding(Figs.S2 and S3).This is discussed in more detail in the Sections S1.1S1.5 in the Supplementary data.Cells with surface displayed LBDs were applied for a flow cytometry-based binding assay to evaluate fluorescent TCN-201 derivatives[4](Figs.1B,S4,and S5).More detailed information about the structural prerequisites of TCN-201 derivatives for interacting with the modulatory binding site and synthesis of fluorescent derivatives is given in the Sections S1.6 and S1.7 in the Supplementary data.展开更多
基金supported by the Research Training Group“Chemical biology of ion channels(Chembion)”,funded by the German Research Foundation(DFG),Germany(Project No:404595355,GRK:2515).
文摘Autodisplay of N-methyl-D-aspartate(NMDA)receptor[1]ligand binding domains(LBDs)was done by inserting the coding sequence for each LBD[2]into a plasmid for maximized autotransporter-mediated expression(pMATE)between the coding sequences of signal peptide and translocator domain[3](Figs.1A and S1).Surface display and functionality of LBDs was confirmed by protease accessibility and radioligand binding(Figs.S2 and S3).This is discussed in more detail in the Sections S1.1S1.5 in the Supplementary data.Cells with surface displayed LBDs were applied for a flow cytometry-based binding assay to evaluate fluorescent TCN-201 derivatives[4](Figs.1B,S4,and S5).More detailed information about the structural prerequisites of TCN-201 derivatives for interacting with the modulatory binding site and synthesis of fluorescent derivatives is given in the Sections S1.6 and S1.7 in the Supplementary data.
