The major objective of this research is to investigate the biosynthesis of the asparagine-linked oligosaccharides of the gonadotropin, the glycoprotein hormones of placental and pituitary origin. Specifically, the calf pituitary system will be used and the focus of attention will center upon bovine luteinizing hormone, which, unlike any other glycoprotein, has N-acetylgalactosamine as an integral part of the asparagine-linked carbohydrate unit. The first objective of the proposal related to the characterization of the lipid-linked intermediates in the biosynthesis of calf pituitary glycoproteins has been achieved. Secondly, incorporation of N-acetylagalactosamine in the oligosaccharide chain of bLH will be investigated. Third, the role of glucose-containing oligosaccharides as intermediates in the process of asparagne-linked oligosaccharide assembly and attachment will be considered. Fourth, the enzymes involved in the processing, recognition, and synthesis of the inner-core will be isloated and studied. In particular, the enzyme which transfers the lipid-linked oligosaccharide to protein will be the first to be isolated in the pituitary system. This will require the use of suitable nonglycosylated acceptors and appropriate oligosaccharidedolichol phosphate compounds. Fifth, experiments will be conducted which will seek to correlate the physiological effects of such agents as LH-RH, cAMP, steroids, and phosphodiesterase inhibitors with their effects upon the glycosylation reaction. Finally, the complete in vitro synthesis of bLH using mRNA and reticulocyte translation system will be attempted.