A simple method for the purification of dipeptidyl aminopeptidase IV from porcine kidney using HPLC.
Dipeptidyl aminopeptidase IV (E.C. 3.4.14.5) has been purified to homogeneity using "soft-column" chromatography on Sephadex G-200, DEAE-cellulose, gly-pro-AH-Sepharose and finally HPLC on TSK G-3000SW. The purification by HPLC is fast and gives a better yield of pure enzyme than procedures which have been described previously. The enzyme obtained in this manner has a high specific activity (86 U/mg using a saturating level of gly-pro-4-nitroanilide as substrate) and was free of contaminating peptidase activities. Our preparation of DAP IV is suitable for peptide sequencing studies by fast atom bombardment mass spectrometry.
Hutchinson
D W
DW
Department of Chemistry, University of Warwick, Coventry, UK.
Woolfitt
A R
AR
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D018819:Dipeptidyl Peptidase 4; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000032:analysis / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D007668:Kidney / Q000201:enzymology*; D013552:Swine
D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; D018819:Dipeptidyl Peptidase 4