Growth hormone (GH) and a GH antagonist promote GH receptor dimerization and internalization.
It has previously been shown that a human growth hormone (hGH) analog, hGH-G120R, acts as a GH antagonist (Chen, W. Y., Wight, D. C. , Wagner, T. E., and Kopchick, J. J. (1990) Proc. Natl. Acad. Sci. U. S. A. 87, 5061-5065; Chen, W. Y., White, M. E., Wagner, T. E., and Kopchick, J. J. (1991) Endocrinology 129, 1402-1408; Chen, W. Y., Chen, N-Y., Yun, J., Wang, X. Z., Wagner, T. E., and Kopchick, J. J. (1994) J. Biol. Chem. 269, 15892-15897). In this study, we report the ability of hGH and hGH-G120R to be internalized by GH receptor expressing cells. Additionally, results of chemical cross-linking experiments revealed that both native hGH and hGH-G120R form complexes similar in size to that expected for hGH when bound to recombinant hGH-binding protein (bp). The molecular mass of the complex was determined to be approximately 280 kDa which is consistent with multiple receptors interacting with the ligand. The predominant radiolabeled band detected was a complex of approximately 140 kDa which probably represents one GH molecule bound to one GH receptor. The cross-linked complexes were not detected in the presence of excess unlabeled hGH or hGH-G120R and were not observed in cells which do not express detectable levels of GH receptors. Also, GH induced tyrosine phosphorylation of a complex of proteins of approximately 95 kDa in these cells whereas hGH-G120R did not. Thus, we have separated the hGH or hGH-G120R/GHR binding and internalization capabilities from the ability to stimulate tyrosine phosphorylation of intracellular proteins.
Harding
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Edison Biotechnology Institute, Molecular and Cellular Biology Program, Ohio University, Athens, Ohio 45701-2979, USA.
Wang
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X
Okada
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Chen
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WY
Wan
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Kopchick
J J
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D000818:Animals; D001704:Biopolymers; D004705:Endocytosis; D013006:Growth Hormone / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors / Q000494:pharmacology / Q000502:physiology*; D006801:Humans; D007457:Iodine Radioisotopes; D051379:Mice; D010750:Phosphoproteins / Q000096:biosynthesis; D010766:Phosphorylation; D011986:Receptors, Somatotropin / Q000378:metabolism*; D011994:Recombinant Proteins; D014443:Tyrosine / Q000378:metabolism
D001704:Biopolymers; C093172:G120R protein, human; D007457:Iodine Radioisotopes; D010750:Phosphoproteins; D011986:Receptors, Somatotropin; D011994:Recombinant Proteins; D014443:Tyrosine; D013006:Growth Hormone