Isolation and characterization of dipeptidyl aminopeptidase IV from human kidney cortex

Clin Chim Acta. 1989 Jan 13;179(1):61-71. doi: 10.1016/0009-8981(89)90023-5.

Abstract

Intact dipeptidyl aminopeptidase IV (DAP IV) was solubilized by bromelain treatment from human kidney brush border plasma-membranes. Purification of DAP IV was performed by a 3-step method, applying lectin-affinity chromatography on WGA-Sepharose, gel filtration and anion-exchange chromatography. DAP IV from human kidney cortex showed a pH optimum of 8.7 and was totally inhibited by 1 mmol/l Zn2+. Isolated DAP IV revealed a relative molecular mass of 250 kDa as determined by the native-PAGE method and of 220 kDa by the gel filtration method. Analytical isoelectric focussing of DAP IV revealed an isoelectric point of pH 5.3. Ultrastructural analysis of isolated DAP IV fractions, using the negative staining technique, disclosed the presence of numerous globular particles with an average diameter of 5 nm which correspond to the structural substrate of the purified protein.

MeSH terms

  • Dipeptidyl Peptidase 4
  • Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / analysis
  • Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / isolation & purification*
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Hydrogen-Ion Concentration
  • Isoelectric Focusing
  • Kidney Cortex / analysis
  • Kidney Cortex / enzymology*
  • Kidney Cortex / ultrastructure

Substances

  • Dipeptidyl-Peptidases and Tripeptidyl-Peptidases
  • Dipeptidyl Peptidase 4