Isolation, characterization and cDNA sequencing of a Kazal family proteinase inhibitor from seminal plasma of turkey (Meleagris gallopavo)

Comp Biochem Physiol B Biochem Mol Biol. 2008 Jun;150(2):207-15. doi: 10.1016/j.cbpb.2008.03.003. Epub 2008 Mar 13.

Abstract

The turkey reproductive tract and seminal plasma contain a serine proteinase inhibitor that seems to be unique for the reproductive tract. Our experimental objective was to isolate, characterize and cDNA sequence the Kazal family proteinase inhibitor from turkey seminal plasma and testis. Seminal plasma contains two forms of a Kazal family inhibitor: virgin (Ia) represented by an inhibitor of moderate electrophoretic migration rate (present also in the testis) and modified (Ib, a split peptide bond) represented by an inhibitor with a fast migration rate. The inhibitor from the seminal plasma was purified by affinity, ion-exchange and reverse phase chromatography. The testis inhibitor was purified by affinity and ion-exchange chromatography. N-terminal Edman sequencing of the two seminal plasma inhibitors and testis inhibitor were identical. This sequence was used to construct primers and obtain a cDNA sequence from the testis. Analysis of a cDNA sequence indicated that turkey proteinase inhibitor belongs to Kazal family inhibitors (pancreatic secretory trypsin inhibitors, mammalian acrosin inhibitors) and caltrin. The turkey seminal plasma Kazal inhibitor belongs to low molecular mass inhibitors and is characterized by a high value of the equilibrium association constant for inhibitor/trypsin complexes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Avian Proteins / chemistry*
  • Avian Proteins / genetics
  • Avian Proteins / isolation & purification
  • Base Sequence
  • Cloning, Molecular
  • DNA, Complementary / chemistry
  • Male
  • Molecular Sequence Data
  • Semen / chemistry*
  • Sequence Alignment
  • Testis / chemistry
  • Trypsin Inhibitor, Kazal Pancreatic / chemistry*
  • Trypsin Inhibitor, Kazal Pancreatic / genetics
  • Trypsin Inhibitor, Kazal Pancreatic / isolation & purification
  • Turkeys* / genetics

Substances

  • Avian Proteins
  • DNA, Complementary
  • Trypsin Inhibitor, Kazal Pancreatic