Metabolism of diacylglycerol pyrophosphate by suspension cultured Catharanthus roseus cells identification and characterization of diacylglycerol pyrophosphate phosphatase in plants

Bettina Riedel, Michael Morr, Wen I. Wu, George M. Carman, Josef B. Wissing

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13 Scopus citations

Abstract

Diacylglycerol pyrophosphate (DGPP) is a novel phospholipid found in plants that is the product of the reaction catalyzed by phosphatidate kinase. In this study we examined the metabolism of DGPP in Catharanthus roseus cells. Metabolic labeling studies showed that DGPP was rapidly metabolized to phosphatidate and P(i). The enzyme catalyzing this reaction was named DGPP phosphatase. DGPP phosphatase activity was 20 times more active than phosphatidate kinase activity. Microsomal and soluble isoforms of DGPP phosphatase were found in C. roseus cells. The microsomal enzyme was primarily associated with intracellular membranes. The pH optima of the microsomal and soluble DGPP phosphatases were 4.5 and respectively. Both activities were independent of a divalent cat on requirement and were insensitive to EDTA and N-ethylmaleimide. The DGPP phosphatase activities were inhibited by NaF, MgCl2, MnCl2, and sphingosine. The dependence of the microsomal find soluble DGPP phosphatase activities on DGPP were dose- dependent with maximum activities obtained at. 100 μM (3 mol%)and 600 μM (20 mol%), respectively, using a Triton X-100/DGPP mixed micellar substrate. Pyrophosphate, p-nitrophenylplhosphate. and adenosine diphosphate (ADP) were not substrates for the DGPP phosphatase enzymes. DGPP phosphatase activity was also found in a wide range of organisms including Escherichia coli. Saccharomyces cerevisiae, rat, pig, and cattle.

Original languageEnglish (US)
Pages (from-to)1-10
Number of pages10
JournalPlant Science
Volume128
Issue number1
DOIs
StatePublished - Sep 30 1997

All Science Journal Classification (ASJC) codes

  • Genetics
  • Agronomy and Crop Science
  • Plant Science

Keywords

  • Catharanthus roseus
  • DGPP phosphatase
  • Suspension cultured plant cells

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