Identification and functional characterization of a presqualene diphosphate phosphatase

Koichi Fukunaga, Makoto Arita, Minoru Takahashi, Andrew J. Morris, Michael Pfeffer, Bruce D. Levy

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Presqualene diphosphate (PSDP) is a bioactive lipid that rapidly remodels to presqualene monophosphate (PSMP) upon cell activation (Levy, B. D., Petasis, N. A., and Serhan, C. N. (1997) Nature 389, 985-990). Here, we have identified and characterized a phosphatase that converts PSDP to PSMP. Unlike the related polyisoprenyl phosphate farnesyl diphosphate (FDP), PSDP was not a substrate for type 2 lipid phosphate phosphohydrolases. PSDP phosphatase activity was identified in activated human neutrophil (PMN) extracts and partially purified in the presence of Nonidet P-40 with gel filtration and anion exchange chromatography. Peptide sequencing of a candidate phosphatase was consistent with phosphatidic acid phosphatase domain containing 2 (PPAPDC2), an uncharacterized protein that contains a lipid phosphate phosphohydrolase consensus motif. Recombinant PPAPDC2 displayed diphosphate phosphatase activity with a substrate preference for PSDP > FDP > phosphatidic acid. PPAPDC2 activity was independent of Mg2+ and optimal at pH 7.0 to 8.0. Incubation of [14C]FDP with recombinant human squalene synthase led to [14C]PSDP and [14C]squalene formation, and in the presence of PPAPDC2, [14C]PSMP was generated from [ 14C]PSDP. PPAPDC2 mRNA was detected in human PMN, and is widely expressed in human tissues. Together, these findings indicate that PPAPDC2 in human PMN is the first lipid phosphate phosphohydrolase identified for PSDP. Regulation of this activity of the enzyme may have important roles for PMN activation in innate immunity.

Original languageEnglish
Pages (from-to)9490-9497
Number of pages8
JournalJournal of Biological Chemistry
Volume281
Issue number14
DOIs
Publication statusPublished - 2006 Apr 7
Externally publishedYes

Fingerprint

Phosphatidate Phosphatase
Phosphoric Monoester Hydrolases
Lipids
Phosphates
Polyisoprenyl Phosphates
Farnesyl-Diphosphate Farnesyltransferase
Chemical activation
Squalene
Phosphatidic Acids
presqualene pyrophosphate
Diphosphates
Substrates
Chromatography
Innate Immunity
Gel Chromatography
Anions
Ion exchange
Neutrophils
Gels
Tissue

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification and functional characterization of a presqualene diphosphate phosphatase. / Fukunaga, Koichi; Arita, Makoto; Takahashi, Minoru; Morris, Andrew J.; Pfeffer, Michael; Levy, Bruce D.

In: Journal of Biological Chemistry, Vol. 281, No. 14, 07.04.2006, p. 9490-9497.

Research output: Contribution to journalArticle

Fukunaga, Koichi ; Arita, Makoto ; Takahashi, Minoru ; Morris, Andrew J. ; Pfeffer, Michael ; Levy, Bruce D. / Identification and functional characterization of a presqualene diphosphate phosphatase. In: Journal of Biological Chemistry. 2006 ; Vol. 281, No. 14. pp. 9490-9497.
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