The Hinge Region of Human Thyroid-Stimulating Hormone (TSH) Receptor Operates as a Tunable Switch between Hormone Binding and Receptor Activation
Figure 5
cAMP production by TSHR truncation or chimeric mutants in response to TSH and MAb 311.62.
A. Schematic representation of hTSHR constructs (not drawn to scale). Soluble plasma membrane preparation from HEK 293 cells, transiently transfected with various TSH receptor constructs were electrophoresed and probed with either (B(i))TSHR HinR specific polyclonal antibody or (B(ii))Anti-His tag monoclonal antibody. A similar full length hTSHR tagged at C-terminal with His-Tag (hTSHRmychis) was created as a positive control. C. HEK293 cells transiently transfected with hTSHR constructs were incubated with hTSH (2 nM) for 15 minutes or with MAb 311.62 IgG or NR-IgG (100 µg/ml) for 1 h at 37°C, and cAMP produced was determined. The results presented are representative of two independent experiments.