Intracellular proteolysis of pancreatic zymogens

Yale J Biol Med. Sep-Oct 1992;65(5):407-20; discussion 437-40.

Abstract

Activation of pancreatic digestive zymogens within the pancreatic acinar cell may be an early event in the development of pancreatitis. To detect such activation, an immunoblot assay has been developed that measures the relative amounts of inactive zymogens and their respective active enzyme forms. Using this assay, high doses of cholecystokinin or carbachol were found to stimulate the intracellular conversion of at least three zymogens (procarboxypeptidase A1, procarboxypeptidase B, and chymotrypsinogen 2) to their active forms. Thus, this conversion may be a generalized phenomenon of pancreatic zymogens. The conversion is detected within ten minutes of treatment and is not associated with changes in acinar cell morphology; it has been predicted that the lysosomal thiol protease, cathepsin B, may initiate this conversion. Small amounts of cathepsin B are found in the secretory pathway, and cathepsin B can activate trypsinogen in vitro; however, exposure of acini to a thiol protease inhibitor (E64) did not block this conversion. Conversion was inhibited by the serine protease inhibitor, benzamidine, and by raising the intracellular pH, using chloroquine or monensin. This limited proteolytic conversion appears to require a low pH compartment and a serine protease activity. After long periods of treatment (60 minutes), the amounts of the active enzyme forms began to decrease; this observation suggested that the active enzyme forms were being degraded. Treatment of acini with E64 reduced this late decrease in active enzyme forms, suggesting that thiol proteases, including lysosomal hydrolases, may be involved in the degradation of the active enzyme forms. These findings indicate that pathways for zymogen activation as well as degradation of active enzyme forms are present within the pancreatic acinar cell.

MeSH terms

  • Animals
  • Benzamidines / pharmacology
  • Biological Transport
  • Carbachol / pharmacology
  • Carboxypeptidase B
  • Carboxypeptidases / metabolism
  • Carboxypeptidases A
  • Cholecystokinin / pharmacology
  • Chymotrypsinogen / metabolism
  • Cysteine Proteinase Inhibitors / pharmacology
  • Enzyme Activation / drug effects
  • Enzyme Precursors / metabolism*
  • Hydrogen-Ion Concentration
  • Leucine / analogs & derivatives
  • Leucine / pharmacology
  • Models, Biological
  • Pancreas / cytology
  • Pancreas / drug effects
  • Pancreas / metabolism*
  • Pancreatitis / etiology
  • Protein Processing, Post-Translational* / drug effects
  • Time Factors
  • Trypsin Inhibitors / pharmacology

Substances

  • Benzamidines
  • Cysteine Proteinase Inhibitors
  • Enzyme Precursors
  • Trypsin Inhibitors
  • Carbachol
  • Cholecystokinin
  • Chymotrypsinogen
  • Carboxypeptidases
  • Carboxypeptidases A
  • Carboxypeptidase B
  • Leucine
  • benzamidine
  • E 64