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Partial purification and characterization of PAF acetylhydrolase in human amniotic fluid

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Abstract

Platelet-activating factor (PAF) acetylhydrolase, which removes the acetyl moiety at thesn-2 position, has been found in human amniotic fluid. We purified this enzyme by ammonium sulfate precipitation, and sequential use of DEAE-Sepharose CL-6B, hydroxyapatite, chelating-Sepharose, and Mono Q column chromatographies. This enzyme exhibited broad pH optima and was unaffected by EDTA. Partially purified enzyme had a molecular weight of approximately 34 kDa on SDS-PAGE. In addition, the enzyme activity was inhibited by either diisopropylfluorophosphate(DFP) orp-bromophenacylbromide (p-BPB), suggesting that this enzyme possesses active serine and histidine residues. The enzyme showed similar activity towards PAF and oxidatively modified phosphatidylcholine, but didn’t hydrolyze phosphatidylcholine or phosphatidylethanolamine with a long chain fatty acyl group atsn-2 position.

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Authors and Affiliations

  1. College of Pharmacy, Yeungnam University, 712-749, Gyongsan, Korea

    So Young Son & Hyeun Wook Chang

  2. National Insititution of Safety Reaserch (NISR), Seoul, Korea

    So Hee Kim

  3. Emory University, Atlanta, Georgia, USA

    Suk Hwan Baek

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  1. So Young Son
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Son, S.Y., Kim, S.H., Baek, S.H. et al. Partial purification and characterization of PAF acetylhydrolase in human amniotic fluid. Arch. Pharm. Res. 20, 218–224 (1997). https://doi.org/10.1007/BF02976148

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