title: Intracellular processing of apolipoprotein J precursor to the mature heterodimer.
abstract: Circulating apolipoprotein J (apoJ) is a 70 kDa glycoprotein comprised of disulfide-linked alpha and beta subunits derived from a single precursor. Post-translational modifications that occur prior to apoJ secretion were assessed, with specific focus on carbohydrate type, the timing of proteolytic cleavage, and the importance of glycosylation on the cleavage and secretion processes. ApoJ was initially resolved as a single chain, intracellular precursor of 58 kDa which contained N-linked oligosaccharide but no O-linked oligosaccharide. The precursor was converted to an intracellular 70 kDa glycoprotein, which became the major intracellular form of apoJ prior to secretion. Maturation of the 58 kDa precursor involved conversion of high-mannose carbohydrate to complex-type carbohydrate containing sialic acid, as well as intracellular cleavage to yield alpha and beta subunits. This cleavage event occurred at a late stage of carbohydrate modification, most likely in the trans-Golgi or a post-Golgi compartment. The maturation and secretion of apoJ occurred rapidly, with a half-time of 30-35 min. Tunicamycin treatment of cells resulted in an unglycosylated doublet comprised of one single chain and one cleaved form of apoJ. The unglycosylated apoJ species were secreted rapidly with a half-time of 20 min. Both cleavage and secretion were independent of glycosylation.
authors.affiliation: Developmental Biology Program, University of Cincinnati, College of Medicine, OH 45267-0575.
authors.forename: B F
authors.initials: BF
authors.lastname: Burkey
authors.forename: H V
authors.initials: HV
authors.lastname: deSilva
authors.forename: J A
authors.initials: JA
authors.lastname: Harmony
mesh_terms: D001053:Apolipoproteins / Q000235:genetics / Q000378:metabolism*; D002236:Carbohydrate Conformation; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D002460:Cell Line; D051152:Clusterin; D006023:Glycoproteins / Q000096:biosynthesis / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D006868:Hydrolysis; D008075:Lipoproteins, HDL / Q000235:genetics / Q000378:metabolism*; D008113:Liver Neoplasms / Q000378:metabolism; D018832:Molecular Chaperones; D011487:Protein Conformation; D011498:Protein Precursors / Q000096:biosynthesis / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional; D014407:Tumor Cells, Cultured; D014415:Tunicamycin / Q000494:pharmacology
chemical_list: D001053:Apolipoproteins; C495242:CLU protein, human; D051152:Clusterin; D006023:Glycoproteins; D008075:Lipoproteins, HDL; D018832:Molecular Chaperones; D011498:Protein Precursors; D014415:Tunicamycin