Abstract
Tissue fragments and isolated acinar cells from adult rat pancreas rapidly incorporated inorganic 35SO 4 2− into macromolecules. Secretion of labeled macromolecules could be stimulated by the secretagogue carbamylcholine chloride. The principal sulfated product synthesized by isolated acinar cells was purified by diethylaminoethyl cellulose chromatography and was identified as heparan sulfate by digestion with purified heparitinase and by deaminative cleavage with nitrous acid. The results of subcellular fractionation of pancreas tissue labeled in vitro with 35SO 4 2− showed that sulfate-containing macromolecules were present both in soluble fractions and as tightly bound membrane components. Zymogen granules contained 14% of the total radioactivity incorporated; zymogen granule membranes, 3%; the soluble zymogen granule contents, 7%; microsomes, 30%; mitochondria, 14%; and the postmicrosomal supernate, 25%. Purified 35SO 4 2−-labeled zymogen granule membranes had the highest specific radioactivity of any fraction examined. Degradative analysis with nitrous acid indicated that the major sulfated product present in all of the subcellular fractions was heparan sulfate.
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