A combination of unnatural phosphatidyl acceptor and tandem electrospray ionization mass spectrometry for tracing phospholipase d activity

Keiichi Oda, Megumi Imura, Yoshimi Ueda, Miho Hosokawa, Michiyo Kobayashi, Junko Matsubara, Mizuho Sato, Naokazu Kumura, Minoru Izumi, Shuhei Nakajima, Tsuyoshi Sugio, Naomichi Baba

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Abstract

Phospholipase D (PLD) is a biocatalyst in the synthesis of bioactive compounds and a key enzyme in a variety of biological signal transductions. A combination of unnatural phosphatidyl acceptor, N,N,N-triethyl-N-2- hydroxyethylammonium bromide 6, as a substrate for PLD, and tandem electrospray ionization mass spectrometry (ESI MS) was found to provide information as to whether a given phospholipid serves as a substrate for the PLD-catalyzed reaction. Thus 2-(13′-hydroperoxy-octadecadienoyl)-1- palmitoylglycerophosphocholine 1, and its degradation products 2-(130-oxo-octadecadienoyl)-1-palmitoylglycerophosphocholine 9 and 2-(13′-hydroxy-octadecadienoyl)-1-palmitoylglycerophosphocholine 11, in a mixture were found to be a substrate of the PLD-catalyzed transphosphatidylation. The sensitivity of this method was exemplified by the observation that PLD activity in cabbage leaves was detected using a small amount of crude crushed leaves with little pretreatment. This simple method can be used in screening for PLD activity and searching for inhibitors of the enzyme from various natural sources.

Original languageEnglish
Pages (from-to)1233-1237
Number of pages5
JournalBioscience, Biotechnology and Biochemistry
Volume73
Issue number5
DOIs
Publication statusPublished - 2009

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Keywords

  • ESI MS
  • Hydroperoxide
  • Phosphatidylcholine
  • Phospholipase D

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Applied Microbiology and Biotechnology
  • Analytical Chemistry
  • Organic Chemistry

Cite this

Oda, K., Imura, M., Ueda, Y., Hosokawa, M., Kobayashi, M., Matsubara, J., Sato, M., Kumura, N., Izumi, M., Nakajima, S., Sugio, T., & Baba, N. (2009). A combination of unnatural phosphatidyl acceptor and tandem electrospray ionization mass spectrometry for tracing phospholipase d activity. Bioscience, Biotechnology and Biochemistry, 73(5), 1233-1237. https://doi.org/10.1271/bbb.90093