Complex Formation of Human Proelastases with Procarboxypeptidases A1 and A2

J Biol Chem. 2016 Aug 19;291(34):17706-16. doi: 10.1074/jbc.M116.743237. Epub 2016 Jun 29.

Abstract

The pancreas secretes digestive proenzymes typically in their monomeric form. A notable exception is the ternary complex formed by proproteinase E, chymotrypsinogen C, and procarboxypeptidase A (proCPA) in cattle and other ruminants. In the human and pig pancreas binary complexes of proCPA with proelastases were found. To characterize complex formation among human pancreatic protease zymogens in a systematic manner, we performed binding experiments using recombinant proelastases CELA2A, CELA3A, and CELA3B; chymotrypsinogens CTRB1, CTRB2, CTRC, and CTRL1; and procarboxypeptidases CPA1, CPA2, and CPB1. We found that proCELA3B bound not only to proCPA1 (KD 43 nm) but even more tightly to proCPA2 (KD 18 nm), whereas proCELA2A bound weakly to proCPA1 only (KD 152 nm). Surprisingly, proCELA3A, which shares 92% identity with proCELA3B, did not form stable complexes due to the evolutionary replacement of Ala(241) with Gly. The polymorphic nature of position 241 in both CELA3A (∼4% Ala(241) alleles) and CELA3B (∼2% Gly(241) alleles) points to individual variations in complex formation. The functional effect of complex formation was delayed procarboxypeptidase activation due to increased affinity of the inhibitory activation peptide, whereas proelastase activation was unchanged. We conclude that complex formation among human pancreatic protease zymogens is limited to a subset of proelastases and procarboxypeptidases. Complex formation stabilizes the inhibitory activation peptide of procarboxypeptidases and thereby increases zymogen stability and controls activation.

Keywords: complex; elastase; metalloprotease; pancreas; pancreatitis; proteolytic enzyme; serine protease; zymogen activation.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Substitution
  • Animals
  • Carboxypeptidase B / genetics
  • Carboxypeptidase B / metabolism
  • Carboxypeptidases A / genetics
  • Carboxypeptidases A / metabolism*
  • Cattle
  • Cell Line
  • Enzyme Activation / physiology
  • Enzyme Precursors / genetics
  • Enzyme Precursors / metabolism*
  • Humans
  • Mutation, Missense
  • Pancreatic Elastase / genetics
  • Pancreatic Elastase / metabolism*
  • Swine

Substances

  • CPB1 protein, human
  • Enzyme Precursors
  • Carboxypeptidases A
  • Carboxypeptidase B
  • Pancreatic Elastase

Associated data

  • PDB/1PYT