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NAE Metabolism

N-Acylphosphotidylethanolamine (NAPE) is an unusual phospholipid with a third acyl moiety linked to the ethanolamine head group of phosphatidylethanolamine (PE). NAPE is present in membranes of plants, animals, and some microorganisms, where it is normally present at only a few percent of the total phospholipid fraction.  The activation of N-acylphosphatidylethanolamine (NAPE) metabolism in plants appears to be associated mostly with cellular stresses. In response to pathogen elicitors, NAPE is hydrolyzed by phospholipase-D (PLD), and corresponding medium-chain, saturated N-acylethanolamines (NAEs) are released by plant cells where they act as lipid mediators to modulate ion flux and activate defense gene expression.  In desiccated seeds of higher plants, long-chain, saturated and unsaturated NAEs are prevalent, but are rapidly metabolized during the first few hours of imbibition, a period of substantial osmotic stress. NAPE synthesis is increased in seeds during this same period of rapid rehydration. A membrane-bound enzyme designated NAPE synthase has been purified from imbibed cottonseeds and its unusual biochemical properties suggest that it may scavenge free fatty acids in vivo.  This feature of NAPE metabolism may be unique to higher plants a may be a mechanism for the rapid recycling of fatty acids back into membrane-associated NAPE (below).  Altogether, increasing evidence indicates that NAPE metabolism in plants shares functional similarities with NAPE metabolism in animal systems, including signal transduction and cellular protection.   In particular, the emerging role of released NAEs as lipid mediators in plant defense signaling represents an intriguing parallel to “endocannabinoid signaling” in several mammalian cell types. In support of this concept, an ortholog of the mammalian fatty acid amide hydrolase was identified in Arabidopsis and other plant species indicating conservation of some of the machinery for NAE metabolism between plants and animals.

 

 

 

 

 

 

Proposed Scheme for the metabolism of N-acylethanolamines (NAEs) in Plants and Animals

nae metabolism scheme
In both plants and animals NAEs are either metabolized to NAE oxylipins (lipoxygenase - LOX) or hydrolyzed to free fatty acids (FFAs) and ethanolamine (fatty acid amide hydrolase - FAAH). FFAs are formed by NAE (FAAH) or or phospholipid (PLA) hydrolysis. NAPE is formed from FFA via NAPE synthase in plants and by coordinated acyltransferase-transacylase in animals.To complete the cycle, NAPE can form NAE through phosphatidyltransferase activity. Differences and commonalities in plant and animal systems are color-coded above.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Fatty Acid Amide Hydrolase (FAAH)

 

Schematic drawing of FAAH domains from information obtained from alignment and prediction programs. The amidase region has a 3 part domain structure which is highlighted in red, green, and blue. Mouse rollover of these areas highlight the corresponding region in the structural model of the FAAH protein.

 

FAAH Knockouts and Overexpressors

 

FAAH Knockouts
FAAH ko plants with nae 12:0
NAE12:0 induced a dose-dependent reduction in seedling development in Arabidopsis wild type as indicated by reduced cotyledon area. The effects of NAE12:0 on Arabidopsis cotyledon area was more pronounced in AtFAAH knockouts. Wild-type seedlings are able to recover from exogenous NAE12:0 2 weeks after germination, whereas AtFAAH knockouts remain severely stunted.
FAAH Overexpressors
FAAH OE plants with nae 12:0
  Three independent AtFAAH overexpressors were able to sustain hypocotyl expansion despite elevated levels of exogenous NAE12:0. (D) Extended exposure to 500µM NAE12:0 is strongly inhibitory to vector control and wild-type seedlings. AtFAAH overexpressors, on the other hand, display robust growth despite the high levels and extended exposure to exogenous NAE12:0.
Wang et al. PNAS | August 8, 2006 | vol. 103 | no. 32 | 12199 PLANT BIOLOGY

 

 

NAE Metabolism Publications

  1. Chapman, K.D. and Moore, T.S.,Jr. 1993. Catalytic properties of a newly-discovered acyltransferase that synthesizes N-acylphosphatidylethanolamine in cottonseed (Gossypium hirsutum L.) microsomes. Plant Physiol. 102(3): 761-769.
  2. Chapman, K.D. and Moore, T.S.,Jr. 1994. Isozymes of cottonseed microsomal N-acylphosphatidylethanolamine synthase: Detergent solubilization and electrophoretic separation of active enzymes with different properties. Biochim. Biophys. Acta, 1211: 29-36.
  3. Cai, S.J., McAndrew, R.S., Leonard, B.P., Chapman, K.D., and Pidgeon, C. 1995. Rapid purification of cotton seed membrane-bound N-acylphosphatidylethanolamine synthase by Immobilized Artificial Membrane (IAM) chromatography. J. Chromatography A, 696: 49-62.  
  4. McAndrew, R.S., Leonard, B.P., and Chapman, K.D. 1995. Photoaffinity labeling of cottonseed microsomal N-acylphosphatidylethanolamine synthase protein with a substrate analogue, 12-[(4-azidosalicyl)amino]dodecanoic acid. Biochim. Biophys. Acta, 1256: 310-318.  
  5. Chapman, K.D., Lin, I., and Desouza, A.D. 1995. Metabolism of cottonseed microsomal N-acylphosphatidylethanolamine. Arch. Biochem. Biophys., 318: 401-407.  
  6. Chapman, K.D., Jackson, A.C., Moreau, R.A., and Tripathy, S. 1995. Increased N-acylphosphatidylethanolamine biosynthesis in elicitor-treated tobacco cells. Physiologia Plantarum, 95:120-126.
  7. McAndrew, R.S., and Chapman, K.D. 1998. Enzymology of cottonseed N-acylphosphatidylethanolamine synthase: kinetic properties and mechanism-based inactivation Biochimica Biophysica Acta 1390: 21-36.
  8. Shrestha, R., Noordemeer, M., Van der Stelt, M., Veldink, G.A., and Chapman, K.D. (2002) N-Acylethanolamines are metabolized by lipoxygenase and amidohydrolase in competing pathways during cotton (Gossypium hirsutum, L.) seed imbibition. Plant Physiology 130: 391-401.
  9. Shrestha, R., Dixon, R.A., Chapman, K.D. (2003) Molecular Identification of a Functional Homologue of the Mammalian Fatty Acid Amide Hydrolase in Arabidopsis thaliana. Journal of Biological Chemistry 278(37): 34990-34997.
  10. Chapman, K.D. (2004) Occurrence, metabolism and prospective functions of N-acylethanolamines in plants. Progress in Lipid Research 43: 302-327.
  11. Shrestha, R., Kim, S.-C., Dyer, J., Dixon, R.A., Chapman, K.D. (2006) Plant Fatty Acid (Ethanol) Amide Hydrolases. Biochimica et Biophysica Acta (BBA)- Molecular and Cell Biology of Lipids, 1761(3):324-334 and on-line-doi:10.1016/j.bbalip.2006.03.004 .
  12. Blancaflor, E.B. and Chapman, K.D. (2006) Similarities between endocannabinoid signaling in animal systems and N-acylethanolamine metabolism in higher plants. In Communication in Plants, F. Baluska, S. Mancuso, and D Volkmann eds., Ch. 14, Springer-Verlag, pp 205-219.
  13. Kilaru A, Blancaflor EB, Venables BJ, Tripathy S, Mysore KS, Chapman KD (2007)The N-acylethanolamine-mediated regulatory pathway in plants. Chem Biodivers 4(8):1933-55. Review.

 

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