Expression of biologically active bovine luteinizing hormone in Chinese hamster ovary cells.
Biologically active bovine luteinizing hormone (LH) has been obtained through expression of the alpha- and LH beta-subunit genes in stably transformed clones of DUXB11, a Chinese hamster ovary cell line deficient in dihydrofolate reductase (DHFR). Expression of alpha-and LH beta-subunit mRNAs of the expected sizes (approximately 910 and 770 nucleotides, respectively) were revealed by blot analysis after electrophoresis of total cellular RNA. Furthermore, presence or absence of the gonadotropin mRNAs in several clonal lines was directly correlated with the appearance of one or both bovine LH subunits in the culture medium. Media from three clones secreting significant immunoreactive levels of both subunits also stimulated the release of progesterone in ovine luteal cells, suggesting that the secreted LH was assembled into a biologically active and glycosylated dimer. Immunoprecipitation and NaDodSO4/PAGE of [35S]methionine-labeled proteins secreted from one of the clones, CHODLH20, further confirmed the presence of an alpha/beta dimer with apparent subunit molecular weights of 20,500 and 16,000, only slightly higher than those of pituitary alpha and LH beta subunits.
D000818:Animals; D002417:Cattle; D002460:Cell Line; D003338:Corpus Luteum / Q000187:drug effects / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D004274:DNA, Recombinant / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D005822:Genetic Vectors; D007986:Luteinizing Hormone / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D010053:Ovary; D011374:Progesterone / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D012756:Sheep
D004274:DNA, Recombinant; D011994:Recombinant Proteins; D011374:Progesterone; D007986:Luteinizing Hormone