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Title: Posttranslational modification and secretion of vitellogenin in Xenopus laevis

Technical Report ·
OSTI ID:6641621

The thesis represents an attempt to elucidate the intracellular pathway linking the synthesis and secretion of the yolk precursor protein vitellogenin. More specifically, how the hepatocyte attaches both phosphate and carbohydrate to the peptide backbone prior to secretion is considered. A model which summarizes the accumulated data depicting various aspects of these processes is also presented. Pulse-chase experiments demonstrate the subcellular sites of vitellogenin phosphorylation. The results of these experiments indicate that approximately 70% of the phsosphate residues are covalently attached to vitellogenin during its intracellular translocation through the smooth microsomes, while the rough microsomes can account for the remainder of the total incorporated phosphate. The presence of a divalent-cation requiring protein kinase (E.C. 2.7.1,37, ATP:protein phosphotransferase) is demonstrated in the microsomes derived from the liver of estrogenized female Xenopus laevis. We present data showing that this protein kinase is responsible for phosphorylating hepatic precursors to serum vitellogenin in vivo. Pulse-chase experiments measuring the rates of incorporation of radiolabeled glucosamine and galactose into intracellular vitellogenin show that glycosylation of this multicomponent protein occurs in a Golgi-enriched fraction isolated from homogenized liver slices. The results indicate that the oligosaccharide component of vitellogenin in Xenopus is a complex type of carbohydrate unit which is linked via an N-glycosidic bond between an asparagine residue and N-acetylglucosamine. With respect to subcellular localization of glycoprotein assembly in Xenopus liver, there is a significant departure from currently accepted models of glycoprotein synthesis.

Research Organization:
Tennessee Univ., Knoxville (USA)
DOE Contract Number:
AC05-76OR00033
OSTI ID:
6641621
Report Number(s):
DOE/NBM-1047; ON: DE83003002
Resource Relation:
Other Information: Thesis
Country of Publication:
United States
Language:
English