Purification and characterization of nuclear alkaline phospholipase A 2 in rat ascites hepatoma cells

The alkaline phospholipase A 2 (PLA 2) 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 PLA 2 monoclonal antibody. From these two columns, the alkaline PLA 2 was eluted in parallel with a...

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Bibliographic Details
Published in:FEBS letters 1996-09, Vol.394 (1), p.55-60
Main Authors: Oishi, Takashi, Tamiya-Koizumi, Keiko, Kudo, Ichiro, Iino, Satoshi, Takagi, Kenzo, Yoshida, Shonen
Format: Article
Language:English
Subjects:
2-ME, 2-mercaptoethanol
[ 14C]arachidonyl PE, 1-acyl-2-[1- 14C]arachidonyl phosphatidylethanolamine
Group II
Nucleus
PC, phosphatidylcholine
PE, phosphatidylethanolamine
Phospholipase A 2
PI, phosphatidylinositol
PLA 2, phospholipase A 2
PMSF, phenylmethylsulfonyl fluoride
PS, phosphatidylserine
Purification
Rat hepatoma
SDSPAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis
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Summary:The alkaline phospholipase A 2 (PLA 2) 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 PLA 2 monoclonal antibody. From these two columns, the alkaline PLA 2 was eluted in parallel with a 17-kDa protein which is reactive to another antigroup II PLA 2 polyclonal antibody. Approximately 80% of nuclear PLA 2 was inhibited by this antibody. The alkaline PLA 2 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 PLA 2 antiserum.
ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(96)00929-5