Purification and characterization of human pancreatic phospholipase A2.
We purified human pancreatic phospholipase A2 from postmortem pancreatic tissue by elution of the semi-purified enzyme on CM-Sephadex C-25 with a linear NaCl gradient at pH 6.0. The enzyme appeared as a single polypeptide chain with an isoelectric point of 9.2 +/- 0.1. The relative molecular mass of the enzyme was estimated to be 15 800 +/- 1000 by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. The enzyme is resistant to heating and to a 25 g/L concentration of sodium dodecyl sulfate. It is inhibited by Ca2+ ions in the presence of ovolecithin and deoxycholate. By immunohistochemical methods we showed the enzyme to be localized in the apical zymogen granule portion of pancreatic acinar cells.
D002102:Cadaver; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007124:Immunoenzyme Techniques; D007525:Isoelectric Focusing; D008970:Molecular Weight; D010084:Oxidation-Reduction; D010179:Pancreas / Q000032:analysis / Q000201:enzymology*; D010740:Phospholipases / Q000302:isolation & purification*; D010741:Phospholipases A / Q000302:isolation & purification*; D054467:Phospholipases A2
D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2