| Products/Services Used | Details | Operation |
|---|---|---|
| Antibody Modification and Purification Services | The supernatant was collected by ultracentrifugation at 200,000× g for 45 min and then incubated with G1 anti-Flag affinity resin (Genscript) for 1 h at 4 °C. After incubation, the resin was loaded into a plastic gravity flow column and washed with 30 column volumes of washing buffer (20 mM HEPES, pH 7.4, 500 mM NaCl, 0.005% (w/v) LMNG, 0.001% (w/v) CHS, 5 μM NTS8–13, 100 μM TCEP, 2 mM CaCl2). After incubation, the phosphorylated NTSR1 was purified by reverse G1 anti-Flag affinity resin (Genscript) to remove unligated NTSR1, then concentrated for subsequent complex assembly. | Get A Quote |
| Proteins, Expression, Isolation and Analysis | Get A Quote |
Biased allosteric modulators (BAMs) of G protein-coupled receptors (GPCRs) have been at the forefront of drug discovery owing to their potential to selectively stimulate therapeutically relevant signaling and avoid on-target side effects. Although structures of GPCRs in complex with G protein or GRK in a BAM-bound state have recently been resolved, revealing that BAM can induce biased signaling by directly modulating interactions between GPCRs and these two transducers, no BAM-bound GPCR-arrestin complex structure has yet been determined, limiting our understanding of the full pharmacological profile of BAMs. Herein, we developed a chemical protein synthesis strategy to generate neurotensin receptor 1 (NTSR1) w... More
