Human growth hormone-variant demonstrates a receptor binding profile distinct from that of normal pituitary growth hormone.
We have recently established that the human growth hormone-variant (hGH-V) gene is functional in vivo by documenting its expression in the placenta. We have subsequently generated transformed murine cell lines stably expressing the genes for normal pituitary growth hormone (hGH-N), hGH-V, and each of two chimeric genes generated by exon 3 exchanges, hGH-NV3 and hGH-VN3. In the present study, we utilize these cell lines as sources of hormone to characterize and compare the receptor binding profiles of hGH-N with hGH-V. hGH-V was found to displace 125I-ovine prolactin bound to rat liver microsomes (lactogen binding) and to displace 125I-hGH bound to rabbit liver microsomes (somatogen binding). Therefore, hGH-V would be predicted to display both somatogenic and lactogenic bioactivity, a dual specificity previously thought to be unique to hGH-N. The concentrations of hormone necessary to displace 50% (IC50) of the 125I-hGH from somatogen receptors and 125I-ovine prolactin from lactogen receptors was expressed as a ratio, IC50 somatogen: IC50 lactogen, for each hormone tested. A 7.4-fold difference in this ratio was observed for hGH-N compared to hGH-V, suggesting significantly greater selectivity by hGH-V in binding to the somatogen receptor. The intermediate binding ratios of the hGH-NV3 and hGH-VN3 chimeric proteins confirmed the distinct receptor binding profiles of the two parent hormones and served to identify three amino acids of potential importance in defining their respective receptor binding specificities.
Ray
J
J
Howard Hughes Medical Institute, University of Pennsylvania, Philadelphia 19104-6145.
Okamura
H
H
Kelly
P A
PA
Cooke
N E
NE
Liebhaber
S A
SA
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D002460:Cell Line; D004274:DNA, Recombinant; D014644:Genetic Variation; D013006:Growth Hormone / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008668:Metallothionein / Q000235:genetics; D051379:Mice; D008862:Microsomes, Liver / Q000378:metabolism; D008969:Molecular Sequence Data; D011388:Prolactin / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011817:Rabbits; D051381:Rats; D011981:Receptors, Prolactin / Q000378:metabolism; D011986:Receptors, Somatotropin / Q000378:metabolism*; D014162:Transfection
D004274:DNA, Recombinant; D011981:Receptors, Prolactin; D011986:Receptors, Somatotropin; D011388:Prolactin; D013006:Growth Hormone; D008668:Metallothionein