Evidence in vivo of asynchronous intracellular transport of rat pancreatic secretory proteins.

Labeled proteins which appeared in pancreatic juice after the intravenous (i.v.) injection of [35S]methionine into conscious rats with chronic pancreatic duct fistulae were separated by gel electrophoresis and measured by determination of the radioactivity of each of the separated bands. Radioactivity appeared in the secreted proteins 20 min after injection of the label. In the subsequent 10 min, 6.44% of the radioactivity was found in trypsinogen, whereas 100 min later only 3.4% of the radioactivity was associated with this enzyme. The values at 10 and 100 min for amylase were 10.85% and 21%, respectively, showing an earlier appearance of labeled trypsinogen than of amylase. Chymotrypsinogen behaved similarly to trypsinogen. Early secretion of labeled proteases was also demonstrated by separation of pancreatic proteins by two-dimensional gel electrophoresis followed by fluorography. In pancreatic duct cannulated rats, zymogen granules were prepared 30 and 60 min after injection of the labeled methionine. Determination of the radioactivity of the individual proteins demonstrated a similar time course of the labeling pattern in the zymogen granule fraction to that in pancreatic juice. The results of the experiments suggest an asynchronous secretion of newly synthesized rat pancreatic proteins.