Comparative analysis of the sequences and three-dimensional models of human procarboxypeptidases A1, A2 and B

A full-length cDNA clone for preprocarboxypeptidase B from human pancreas has been isolated and sequenced. The open reading frame is 1 254 bp in length, encoding a protein of 417 amino acids that includes a leader signal peptide of 15 amino acids and a 95-amino acid-long pro-segment. It contains two differences when compared to the sequence reported for pancreas-specific protein, a human serum marker for acute pancreatitis identified as procarboxypeptidase B. The main difference is a previously unreported Cys at position 138, which is needed for the formation of one of the three disulphide bridges. Sequence alignments between human procarboxypeptidases Al, A2 and B and other known forms show that the most conserved region is the enzyme moiety followed by the globular domain of the pro-segment. The maximum variability is found in the connecting region between moieties. The known three-dimensional structures of procarboxypeptidases from bovine and porcine species have been used to model all three human procarboxypeptidases and also to estimate the interaction energies between the different parts of the molecules, in an attempt to gain insight into the structural features responsible for the differences observed in the functionality of the proenzymes, particularly in their proteolytic activation pathways. Taken together, the results obtained confirm that the main determinant for the rate and mode of activation of procarboxypeptidases is the strength of the interaction between the enzyme and the globular domain of the pro-segment, the connecting segment playing a complementary role.