Intracellular transport and tyrosine sulfation of procollagens V.
Several tyrosine residues of the extracellular p-collagens V and collagens V are sulfated [Fessler, L. I., Brosh, S., Chapin, S. and Fessler, J. H. (1986) J. Biol. Chem. 261, 5034-5040]. Here, the sulfation of their intracellular precursors, the procollagens V, was studied. A Golgi-enriched subcellular fraction of chick embryo tendon catalyzed the sulfation of tyrosine residues in both endogenous and added, unsulfated procollagens V with the sulfate donor 3'-phosphoadenosine 5'-[35S]phosphosulfate. Intracellular tyrosine sulfation of procollagen V occurred at a point distal to the cis Golgi compartment as judged by change of the N-linked carbohydrate of procollagen V from being endoglycosidase-H-sensitive to being resistant. The time course of the intracellular modifications of procollagen V was determined by incubating tendons with 3H-labeled amino acids and with [35S]sulfate. The pro alpha(V) chains were synthesised in about 10 min and then assembled into unsulfated procollagen V molecules. Tyrosine sulfation occurred 50 min after completion of polypeptide synthesis and the molecules were successively sulfated in the order in which they had been synthesized. The antimicrotubular drug Nocodazole, which disrupts the spatial organization of the Golgi, decreased the time interval between synthesis of procollagens V and sulfation. The sulfated procollagens V were soon secreted and cut to sulfated p-collagens V. Sulfated pro alpha 1(V) chains were cleaved faster than sulfated pro alpha 1'(V) chains. The relationship of sequential protein modification to spatial cellular organization is discussed.
D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology; D001692:Biological Transport; D002642:Chick Embryo; D003520:Cyclophosphamide / Q000494:pharmacology; D015739:Nocodazole; D011347:Procollagen / Q000378:metabolism*; D013431:Sulfates / Q000378:metabolism*; D013462:Sulfur Radioisotopes; D013710:Tendons / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism*
D001562:Benzimidazoles; D011347:Procollagen; D013431:Sulfates; D013462:Sulfur Radioisotopes; D014443:Tyrosine; D003520:Cyclophosphamide; D015739:Nocodazole