Inhibition of lactogenic activities of ovine prolactin and human growth hormone (hGH) by a novel form of a modified recombinant hGH.
A recombinant analog of human GH (hGH) lacking 13 amino acids at the amino-terminus (Met14hGH) inhibited the hGH- or ovine PRL (oPRL)-stimulated proliferation of Nb2 lymphoma cells and bovine PRL-stimulated fat synthesis and alpha-lactalbumin secretion in explants from bovine lactating mammary gland. The inhibition was competitive in nature, and in Nb2 cells could be abolished by an excess of hGH or oPRL. Inhibition of oPRL-stimulated proliferation of Nb2 cells by Met14hGH could also be specifically abolished by anti-hGH monoclonal antibodies. Met14hGH had no growth-stimulating activity in Nb2 cells and was not cytotoxic. It also did not affect glucose uptake by the mammary gland explants. Met14hGH competed with [125I]hGH for binding to intact Nb2 cells, IM-9 lymphocytes, solubilized microsomal fraction from lactating bovine mammary gland, and microsomal fraction from the liver of female virgin rats, but its affinity for those receptors was 2 orders of magnitude lower than the affinity of hGH. Since Met14hGH used in most experiments contained about 25% impurities and degradation products, a small amount of it was further purified by immunoaffinity chromatography. Two purified fractions, one consisting of a single 20K protein and the other accompanied by a small amount of 25K protein, were obtained. Both fractions exhibited increased inhibition of hGH- or oPRL-stimulated proliferation of Nb2 cells, thus indicating that the inhibitory activity results from the intact Met14hGH molecule. To the best of our knowledge, this is the first report describing the inhibition of lactogenic hormone activities by a modified hGH.
Gertler
A
A
Shamay
A
A
Cohen
N
N
Ashkenazi
A
A
Friesen
H G
HG
Levanon
A
A
Gorecki
M
M
Aviv
H
H
Hadary
D
D
Vogel
T
T
D000818:Animals; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002846:Chromatography, Affinity; D046508:Culture Techniques; D000939:Epitopes / Q000276:immunology; D005227:Fatty Acids / Q000378:metabolism; D005260:Female; D013006:Growth Hormone / Q000031:analogs & derivatives* / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D019382:Human Growth Hormone; D007768:Lactalbumin / Q000378:metabolism; D007774:Lactation; D008055:Lipids / Q000096:biosynthesis; D008223:Lymphoma / Q000473:pathology*; D008321:Mammary Glands, Animal / Q000187:drug effects / Q000378:metabolism*; D011247:Pregnancy; D011388:Prolactin / Q000494:pharmacology*; D051381:Rats; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology*
D000939:Epitopes; D005227:Fatty Acids; D008055:Lipids; D011994:Recombinant Proteins; D019382:Human Growth Hormone; D011388:Prolactin; D013006:Growth Hormone; D007768:Lactalbumin