Differential sorting of lutropin and the free alpha-subunit in cultured bovine pituitary cells.
The glycoprotein hormones lutropin (LH) and follitropin (FSH) are both synthesized by gonadotrophs in the anterior pituitary but are stored in separate secretory granules prior to secretion. Despite having highly homologous beta-subunits and alpha-subunits with the identical amino acid sequence, the Asn-linked oligosaccharides on LH terminate with SO4-GalNAc while those on FSH terminate with sialic acid-Gal. In addition to LH and FSH, gonadotrophs secrete uncombined (free) alpha-subunit which bears the same sulfated oligosaccharides as LH. We have examined the synthesis and secretion of LH and free alpha-subunit in primary cultures of bovine pituitary cells in order to determine if the sulfated oligosaccharides have any impact on sorting. Our results show that newly synthesized free alpha-subunit is secreted exclusively via the constitutive pathway with a t1/2 of 1.8 h and is never found in dense-core secretory granules. In contrast, LH dimer is secreted by both the constitutive and the regulated pathways. Constitutive secretion and arrival in a dense secretory granule both occur with t1/2 values of 1-1.5 h for newly synthesized LH. Sulfation occurs immediately prior to arrival of LH in the secretory granule and is followed by a period of 1-1.5 h before the LH-containing granules become sensitive to release by gonadotropin releasing hormone. As a result the t1/2 for LH secretion in the presence of gonadotropin releasing hormone is 3.5 h. Sulfation of the free alpha-subunit oligosaccharides is not, therefore, sufficient to direct the alpha-subunit to secretory granules, and the information required for directing LH to granules must reside either in the beta-subunit or the alpha beta-complex.
Blomquist
J F
JF
Department of Pathology, Washington University Medical School, St. Louis, Missouri 63110.
Baenziger
J U
JU
D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D002478:Cells, Cultured; D015292:Glycoprotein Hormones, alpha Subunit / Q000096:biosynthesis* / Q000302:isolation & purification; D007700:Kinetics; D007986:Luteinizing Hormone / Q000096:biosynthesis* / Q000302:isolation & purification; D046911:Macromolecular Substances; D008715:Methionine / Q000378:metabolism; D008854:Microscopy, Electron; D010903:Pituitary Gland, Anterior / Q000378:metabolism*; D013431:Sulfates / Q000378:metabolism; D013462:Sulfur Radioisotopes
D015292:Glycoprotein Hormones, alpha Subunit; D046911:Macromolecular Substances; D013431:Sulfates; D013462:Sulfur Radioisotopes; D007986:Luteinizing Hormone; D008715:Methionine