Evidence for generation of the growth hormone-binding protein through proteolysis of the growth hormone membrane receptor.

The growth hormone-binding protein (GHBP) which circulates in plasma is a soluble short form of the membrane growth hormone receptor (GHR). In rats and mice, GHR and GHBP originate from two alternatively spliced mRNAs (4.5 and 1.2 kb). In human and rabbit tissues, a single predominant mRNA of 4.5 kb was detected and it was hypothesized that GHBP could be produced by proteolytic cleavage of the GHR. Using gel filtration and HPLC, we have detected a high level of GH binding activity in media of cells transfected with rabbit GHR cDNA. The [125I]hGH-GHBP complex eluted at the same time as the plasma complex and both the binding affinity and specificity of the BP were comparable to that of rabbit plasma. Immunoprecipitation experiments and Western blots confirmed that GHBP in the media of transfected cells was a 55 kDa protein related to the extracellular domain of the GHR. In contrast, no BP was detected in the media of cells transfected with the cDNA encoding the rat GHR. These results strongly suggest that,...