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Fig. 4. Ligand-binding capacities of the mutant receptors. (A) Transiently transfected ts20 cells were pulse-labelled for 10 minutes and chased for 90 or 240 minutes as indicated. Samples were split into equal amounts and immunoprecipitated (IP) with anti-GHR-T or isolated with btGH and streptavidin beads (btGH) and subjected to reducing SDS-PAGE. Receptor species are indicated as in Fig. 3. p, 110-kDa precursor form; m, 130-kDa mature form. Relative molecular mass standards are shown on the right. (B) Quantification of the ligand binding capacity of wild-type and mutated receptors. The amount of radioactivity was determined using ImageQuant (Molecular Dynamics). Precursor form intensities of the wild-type and mutant receptors after btGH pull-down were divided by precursor form intensities of the wild-type and mutant receptors after immunoprecipitation and represented as a percentage. The values represent the mean ± s.d. of at least two different experiments.
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