Cell-free synthesis and processing of the precursors to the subunits of luteinizing hormone.
Polyadenylated RNA prepared from pituitary glands of ovariectomized rats was translated in heterologous cell-free systems derived from wheat germ and rabbit reticulocytes in the absence and in the presence of pancreatic microsomal membranes. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated that the major 14,000-dalton product immunoprecipitated by antisera to the alpha subunit of rat and bovine luteinizing hormone from membrane-free translations was processed to a major 21,000-dalton and a minor 17,000-dalton product in the presence of microsomal membranes. These membrane-dependent products were sensitive to cleavage by the glycosidase endo-beta-N-acetylglucosaminidase H, but the 14,000-dalton product was resistant. The 21,000-dalton product was sequestered within the microsomal vesicles as shown by its resistance to limited proteolysis capable of digesting the 14,000-dalton precursor. Glycosylated products with apparent Mr = 18,000 and 17,000 were identified from membrane-supplemented translations by immunoprecipitation using antisera to the beta subunit of rat luteinizing hormone. The immunological identity of the glycosylated subunits of luteinizing hormone was confirmed by showing competition of the immunoprecipitation reactions with unlabeled subunits.
D000818:Animals; D002369:Castration; D002462:Cell Membrane / Q000378:metabolism; D002474:Cell-Free System; D004285:Dogs; D005260:Female; D007986:Luteinizing Hormone / Q000096:biosynthesis*; D046911:Macromolecular Substances; D008297:Male; D008970:Molecular Weight; D010179:Pancreas / Q000378:metabolism; D010902:Pituitary Gland / Q000378:metabolism*; D010944:Plants / Q000378:metabolism; D011061:Poly A / Q000378:metabolism*; D014176:Protein Biosynthesis; D051381:Rats; D014908:Triticum / Q000378:metabolism
D046911:Macromolecular Substances; D011061:Poly A; D007986:Luteinizing Hormone