Human apolipoprotein A-I. Post-translational modification by fatty acid acylation.
The human apolipoproteins are secretory proteins some of which have been shown to undergo proteolytic processing and post-translational addition of carbohydrate. Apolipoprotein A-I (apo-A-I), the predominant protein associated with high density lipoproteins, undergoes co-translational proteolytic processing as well as post-translational conversion of proapo-A-I to mature apo-A-I following cellular secretion. Utilizing the human hepatoma cell line HEP-G2, we have established that, in addition to proteolytic processing, secreted nascent apo-A-I is acylated with palmitate. Uniformly labeled [14C]palmitate and [1-14C]palmitate were each incorporated into apo-A-I when analyzed by sodium dodecyl sulfate gel electrophoresis and autoradiography. The acylation of apo-A-I with palmitate was confirmed by immunoprecipitation and gas chromatography/mass spectrometry. Hydroxylamine treatment resulted in the deacylation of apo-A-I. Although three of the apo-A-I isoforms analyzed by two-dimensional gel electrophoresis were shown to contain radio-labeled palmitate, 80% of acylated apo-A-I was in the proapolipoprotein A-I isoform. [14C]Oleate was not incorporated in secreted apo-A-I, indicating the specificity of the acylation of apo-A-I. Incubation of [14C] palmitate-acylated apo-A-I in serum and plasma under conditions in which proapo-A-I is proteolytically cleaved to mature apo-A-I did not result in deacylation. These data establish that fatty acid acylation occurs in human secretory proteins in addition to the previously reported acylation of cellular membrane proteins. These results suggest that the covalent linkage of lipids to apolipoproteins may play a critical role in apolipoprotein and lipoprotein metabolism.
D000215:Acylation; D016632:Apolipoprotein A-I; D001054:Apolipoproteins A / Q000096:biosynthesis*; D006528:Carcinoma, Hepatocellular / Q000032:analysis; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005227:Fatty Acids / Q000378:metabolism*; D005470:Fluorometry; D006801:Humans; D007525:Isoelectric Focusing; D008113:Liver Neoplasms / Q000032:analysis; D019308:Palmitic Acid; D010169:Palmitic Acids / Q000378:metabolism; D011499:Protein Processing, Post-Translational
D016632:Apolipoprotein A-I; D001054:Apolipoproteins A; D005227:Fatty Acids; D010169:Palmitic Acids; D019308:Palmitic Acid