Purification of intracellular phospholipase A2 from rat spleen supernatant by reverse-phase high-performance liquid chromatography |
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Authors: | Hiromasa Tojo Takashi Teramoto Toshio Yamano Mitsuhiro Okamoto |
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Institution: | Department of Biochemistry, Osaka University Medical School, Kita-ku, Osaka 530, Japan |
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Abstract: | Intracellular phospholipase A2 was purified to homogenity from rat spleen supernatant by reverse-phase high-performance liquid chromatography with a trifluoroacetic acid-acetonitrile solvent system. The method simplified the purification procedure, which includes three consecutive chromatographic steps. The recovery of the enzyme activity was greater than 70% with an about 23,000-fold purification. The solvent system did not affect the catalytic properties of the enzyme. Phospholipases A2 from rat spleen, human pancreatic juice, and porcine pancreas were eluted in that order from a column of octadecasilyl silica gel in a similar concentration range of acetonitrile. This result suggests that the phospholipases A2 examined have similar hydrophobicities. This method may be applicable to the purification of phospholipases A2 from other sources. |
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Keywords: | phospholipases HPLC proteins hydrophobic chromatography phospholipids |
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