Biosynthesis of complement protein D by HepG2 cells: a comparison of D produced by HepG2 cells, U937 cells and blood monocytes.
The biosynthesis of complement protein D of the alternative pathway by HepG2 cells, a human hepatocyte cell line, was studied and compared to the biosynthesis of D by U937 cells and blood monocytes. Increasing amounts of antigenic D were detected in HepG2 cell culture supernatants by radioimmunoassay. The kinetics of D synthesis and secretion by HepG2 cells was followed in a pulse-chase study using [35S]cysteine. As analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by autoradiography, only a single D band was seen intra- and extracellularly and both forms had the same apparent molecular weight as D synthesized by U937 cells or purified from serum. Treatment of HepG2 and U937 cells with canavanine, an arginine amino acid analog, to inhibit intracellular processing resulted in slight depression of the apparent molecular weight of D synthesized by these cells. D synthesized by blood monocytes had an apparent molecular weight similar to that synthesized by HepG2 and U937 cells, suggesting that these cell lines do not synthesize and process D differently than normal monocytes. The data demonstrate that the hepatocyte is a site of D synthesis and suggest that D is not synthesized as a precursor molecule.
D002172:Canavanine / Q000494:pharmacology; D006528:Carcinoma, Hepatocellular / Q000201:enzymology*; D002460:Cell Line; D003166:Complement Activating Enzymes / Q000096:biosynthesis*; D011416:Complement Factor D / Q000096:biosynthesis* / Q000302:isolation & purification; D003470:Culture Media; D006801:Humans; D007700:Kinetics; D008113:Liver Neoplasms; D008970:Molecular Weight; D009000:Monocytes / Q000201:enzymology*
D003470:Culture Media; D002172:Canavanine; D003166:Complement Activating Enzymes; C493488:CFD protein, human; D011416:Complement Factor D