Stabilization of the C-terminal part of pig and horse colipase by carboxypeptidase and trypsin inhibitors

Eur J Biochem. 1981 Mar 16;115(1):99-105. doi: 10.1111/j.1432-1033.1981.tb06203.x.

Abstract

Pig and horse colipases have been purified by a common procedure using trypsin and carboxypeptidase inhibitors as stabilizers. Two forms of pig colipase were identified: a predominant A1 form with about 103-105 residues, and a minor slightly degraded A2 form in which the last two C-terminal residues, Asp and Ser, were lacking. This type of degradation is considerably slowed down by carboxypeptidase inhibitors. A total of four forms of the horse cofactor were characterized: two (A1 and B1) were probably isocolipases which differed by only a few substitutions. Both contained the same number of residues (about 96), an N-terminal valine and an Arg-Ser-Glu-(Glx)1,2-ArgC-terminal sequence. A2 and B2 were slightly degraded forms probably resulting from tryptic cleavage of the Arg-Ser bond in the above sequence. The presence of methionine in the horse cofactor allowed fragmentation by cyanogen bromide. The C-terminal fragment was composed of 16 or 17 residues and contained no histidine. The single histidine of horse B1 was found in the intermediary fragment between Met-18 and Met-(n-17). These data show that the C-terminal parts of both pig and horse colipases are still more exposed to proteolytic degradations than the N-terminal parts. Preliminary attempts to crystallize B1 were carried out.

MeSH terms

  • Animals
  • Carboxypeptidases / antagonists & inhibitors*
  • Chemical Phenomena
  • Chemistry
  • Colipases / isolation & purification*
  • Crystallization
  • Horses
  • Pancreas / enzymology
  • Proteins / isolation & purification*
  • Swine
  • Trypsin Inhibitors / pharmacology*

Substances

  • Colipases
  • Proteins
  • Trypsin Inhibitors
  • Carboxypeptidases