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Lipidomic analysis of N-acylphosphatidylethanolamine molecular species in Arabidopsis suggests feedback regulation by N-acylethanolamines

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Abstract

N-Acylphosphatidylethanolamine (NAPE) and its hydrolysis product, N-acylethanolamine (NAE), are minor but ubiquitous lipids in multicellular eukaryotes. Various physiological processes are severely affected by altering the expression of fatty acid amide hydrolase (FAAH), an NAE-hydrolyzing enzyme. To determine the effect of altered FAAH activity on NAPE molecular species composition, NAE metabolism, and general membrane lipid metabolism, quantitative profiles of NAPEs, NAEs, galactolipids, and major and minor phospholipids for FAAH mutants of Arabidopsis were determined. The NAPE molecular species content was dramatically affected by reduced FAAH activity and elevated NAE content in faah knockouts, increasing by as much as 36-fold, far more than the NAE content, suggesting negative feedback regulation of phospholipase D-mediated NAPE hydrolysis by NAE. The N-acyl composition of NAPE remained similar to that of NAE, suggesting that the NAPE precursor pool largely determines NAE composition. Exogenous NAE 12:0 treatment elevated endogenous polyunsaturated NAE and NAPE levels in seedlings; NAE levels were increased more in faah knockouts than in wild-type or FAAH overexpressors. Treated seedlings with elevated NAE and NAPE levels showed impaired growth and reduced galactolipid synthesis by the “prokaryotic” (i.e., plastidic), but not the “eukaryotic” (i.e., extraplastidic), pathway. Overall, our data provide new insights into the regulation of NAPE–NAE metabolism and coordination of membrane lipid metabolism and seedling development.

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Abbreviations

ABA:

Abscisic acid

DGDG:

Digalactosyldiacylglycerol

EA:

Ethanolamine

FAAH:

Fatty acid amide hydrolase

FFA:

Free fatty acid

FW:

Fresh weight

KO:

Knock-out

LOX:

Lipoxygenase

MGDG:

Monogalactosyldiacylglycerol

MS:

Mass spectrometry

NAE:

N-acylethanolamine

NAPE:

N-acyl PE

NL:

Neutral loss

OE:

Overexpressor

PA:

Phosphatidic acid

PC:

Phosphatidylcholine

PE:

Phosphatidylethanolamine

PG:

Phosphatidylglycerol

PI:

Phosphatidylinositol

PLD:

Phospholipase D

Pre:

Precursor

PS:

Phosphatidylserine

PU:

Polyunsaturated

SPE:

Solid phase extraction

X:Y:

Designates fatty acid or total acyl carbons:total carbon–carbon double bonds

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Acknowledgments

This work was supported by a grant from the United States Department of Energy, Office of Basic Energy Sciences (BES, grant number DE-FG02-05ER15647) and a seed grant from the University of North Texas to KDC. Method development and equipment acquisition at the Kansas Lipidomics Research Center was funded by National Science Foundation (DBI 0521587; MCB 0455318 and 0920663), Kansas IDeA Networks of Biomedical Research Excellence (INBRE) of the National Institute of Health (P20RR16475) and Kansas State University.

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Author notes

  1. Giorgis Isaac

    Present address: Waters Corporation, 34 Maple Street, Milford, MA, 01757, USA

Authors and Affiliations

  1. Department of Biological Sciences, East Tennessee State University, PO Box 70703, Johnson City, TN, 37614, USA

    Aruna Kilaru

  2. Department of Biological Sciences, Center for Plant Lipid Research, University of North Texas, 1155 Union Circle, #305220, Denton, TX, 76203-5017, USA

    Aruna Kilaru, Barney J. Venables & Kent D. Chapman

  3. Division of Biology, Kansas Lipidomics Research Center, Kansas State University, Manhattan, KS, 66506, USA

    Pamela Tamura, Giorgis Isaac & Ruth Welti

  4. Department of Mathematics and Statistics, East Tennessee State University, Johnson City, TN, 37614, USA

    Edith Seier

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  1. Aruna Kilaru
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Correspondence to Aruna Kilaru or Kent D. Chapman.

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A. Kilaru and P. Tamura contributed equally to the work.

A contribution to the Special Issue on Metabolic Plant Biology.

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Kilaru, A., Tamura, P., Isaac, G. et al. Lipidomic analysis of N-acylphosphatidylethanolamine molecular species in Arabidopsis suggests feedback regulation by N-acylethanolamines. Planta 236, 809–824 (2012). https://doi.org/10.1007/s00425-012-1669-z

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