Use of immobilized adenosine deaminase (EC for the rapid purification of native human CD26/dipeptidyl peptidase IV (EC

J Immunol Methods. 1996 Jan 16;189(1):99-105. doi: 10.1016/0022-1759(95)00239-1.


The leukocyte differentiation antigen CD26 identified as dipeptidyl peptidase IV.(EC, cleaves off N-terminal dipeptides from peptides when a proline or alanine is located at the penultimate position. Seminal plasma and especially prostasomes, prostate-derived organelles which occur freely in seminal plasma, contain high amounts of CD26/dipeptidyl peptidase IV and therefore are suitable sources for the purification of the protein. The use of adenosine deaminase (EC affinity chromatography for its purification is described. CD26/dipeptidyl peptidase IV was purified from human seminal plasma and prostasomes by a two step procedure. Ion exchange chromatography on DEAE-Sepharose, followed by affinity chromatography on adenosine deaminase-Sepharose resulted in the pure, native protein with an overall yield ranging from 35 to 55%. The N-terminal sequence of the amphiphilic enzyme purified from human prostasomes was determined to be Met-Lys-Thr-Pro-Trp-Lys-Val-Leu. The preparation obtained was free of contaminating aminopeptidase activity and proved to be very stable (up to 1 month at 37 degrees C). The calf intestinal adenosine deaminase we used is commercially available and can be employed for the purification of human, bovine and rabbit CD26/dipeptidyl peptidase IV. High affinity binding of porcine dipeptidyl peptidase IV was not observed. The availability of a source with high specific activity and the introduction of adenosine deaminase affinity chromatography permits the rapid purification of milligram quantities of natural mammalian CD26/dipeptidyl peptidase IV.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Deaminase*
  • Amino Acid Sequence
  • Chromatography, Affinity
  • Chromatography, Ion Exchange
  • Dipeptidyl Peptidase 4 / isolation & purification*
  • Enzymes, Immobilized*
  • Humans
  • Male
  • Molecular Sequence Data
  • Organelles / enzymology
  • Organelles / immunology
  • Prostate / enzymology
  • Prostate / immunology
  • Semen / enzymology*
  • Semen / immunology*


  • Enzymes, Immobilized
  • Dipeptidyl Peptidase 4
  • Adenosine Deaminase