Expression, tyrosine sulfation, and secretion of yolk protein 2 of Drosophila melanogaster in mouse fibroblasts.
Tyrosine sulfation is a post-translational modification in the trans Golgi that has been found in all animal species studied. In the preceding paper (Baeuerle, P. A., Lottspeich, F., and Huttner, W. B. (1988) J. Biol. Chem. 263, 14925-14929), we have identified the site of tyrosine sulfation in an insect secretory protein, yolk protein 2 (YP2) of Drosophila melanogaster. In the present report, tyrosine sulfation of this protein was examined after expression in a heterologous mammalian cell system. Mouse fibroblasts, transfected with Drosophila YP2 genomic DNA inserted into the eucaryotic expression vector pSV2, secreted the fly protein in sulfated form. Analyses of Drosophila YP2 produced by the mouse cells showed that the features of sulfation of this protein were identical to those previously determined for YP2 isolated from flies. YP2 secreted from mouse fibroblasts was found to be exclusively sulfated on tyrosine residues. The stoichiometry of tyrosine sulfation was approximately 1 mol of sulfate/mol of YP2. Sulfate was linked to the same tyrosine residue as in YP2 isolated from flies, tyrosine 172. These results show that essential parameters of the tyrosine sulfation reaction are very similar in insects and mammals and thus highly conserved in evolution.
D000818:Animals; D002460:Cell Line; D004274:DNA, Recombinant / Q000378:metabolism; D004331:Drosophila melanogaster / Q000235:genetics*; D004527:Egg Proteins / Q000096:biosynthesis / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D005796:Genes; D015151:Immunoblotting; D007739:L Cells / Q000378:metabolism; D051379:Mice; D010957:Plasmids; D011499:Protein Processing, Post-Translational; D013431:Sulfates / Q000378:metabolism; D013462:Sulfur Radioisotopes; D013464:Sulfuric Acids / Q000378:metabolism; D014162:Transfection; D014443:Tyrosine
D004274:DNA, Recombinant; D004527:Egg Proteins; D013431:Sulfates; D013462:Sulfur Radioisotopes; D013464:Sulfuric Acids; D014443:Tyrosine