Purification and characterization of nuclear alkaline phospholipase A2 in rat ascites hepatoma cells.
The alkaline phospholipase A2 (PLA2) was purified from nuclei of rat ascites hepatoma cells (AH7974) by column chromatography with a Sephacryl S-300 column and an immunoadsorbent using anti-group II PLA2 monoclonal antibody. From these two columns, the alkaline PLA2 was eluted in parallel with a 17-kDa protein which is reactive to another anti-group II PLA2 polyclonal antibody. Approximately 80% of nuclear PLA2 was inhibited by this antibody. The alkaline PLA2 was found in association with the chromatin fraction among subnuclear fractions. By an immunocytochemical staining, the nuclei of AH7974 were stained more strongly than other parts of cells with anti-group II PLA2 antiserum.
Oishi
T
T
Second Department of Internal Medicine, Nagoya University School of Medicine, Japan.
Tamiya-Koizumi
K
K
Kudo
I
I
Iino
S
S
Takagi
K
K
Yoshida
S
S
D000818:Animals; D001201:Ascites; D015153:Blotting, Western; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000201:enzymology*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D007150:Immunohistochemistry; D008099:Liver / Q000201:enzymology; D008114:Liver Neoplasms, Experimental / Q000201:enzymology*; D008970:Molecular Weight; D010741:Phospholipases A / Q000737:chemistry* / Q000302:isolation & purification; D054467:Phospholipases A2; D051381:Rats; D014407:Tumor Cells, Cultured
D010741:Phospholipases A; D054467:Phospholipases A2