Merck

Effects of synthetic alkamides on Arabidopsis fatty acid amide hydrolase activity and plant development.

Phytochemistry (2014-12-11)
Lionel Faure, Ronaldo Cavazos, Bibi Rafeiza Khan, Robby A Petros, Peter Koulen, Elison B Blancaflor, Kent D Chapman
RESUMEN

Alkamides and N-acylethanolamines (NAEs) are bioactive, amide-linked lipids that influence plant development. Alkamides are restricted to several families of higher plants and some fungi, whereas NAEs are widespread signaling molecules in both plants and animals. Fatty acid amide hydrolase (FAAH) has been described as a key contributor to NAE hydrolysis; however, no enzyme has been associated with alkamide degradation in plants. Herein reported is synthesis of 12 compounds structurally similar to a naturally occurring alkamide (N-isobutyl-(2E,6Z,8E)decatrienamide or affinin) with different acyl compositions more similar to plant NAEs and various amino alkyl head groups. These "hybrid" synthetic alkamides were tested for activity toward recombinant Arabidopsis FAAH and for their effects on plant development (i.e., cotyledon expansion and primary root length). A substantial increase in FAAH activity was discovered toward NAEs in vitro in the presence of some of these synthetic alkamides, such as N-ethyllauroylamide (4). This "enhancement" effect was found to be due, at least in part, to relief from product inhibition of FAAH by ethanolamine, and not due to an alteration in the oligomerization state of the FAAH enzyme. For several of these alkamides, an inhibition of seedling growth was observed with greater results in FAAH knockouts and less in FAAH over-expressing plants, suggesting that these alkamides could be hydrolyzed by FAAH in planta. The tight regulation of NAE levels in vivo appears to be important for proper seedling establishment, and as such, some of these synthetic alkamides may be useful pharmacological tools to manipulate the effects of NAEs in situ.

MATERIALES
Referencia del producto
Marca
Descripción del producto

SAFC
Isopropyl β-D-1-thiogalactopyranoside
Sigma-Aldrich
Hexane, Laboratory Reagent, ≥95%
Sigma-Aldrich
Hexane, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥99% (GC)
Sigma-Aldrich
Hexane, ReagentPlus®, ≥99%
Supelco
2,5-Dihydroxybenzoic acid, analytical standard
Sigma-Aldrich
Hexane, suitable for HPLC, ≥97.0% (GC)
Sigma-Aldrich
Hexane, HPLC Plus, for HPLC, GC, and residue analysis, ≥95%
Sigma-Aldrich
Hexane, suitable for HPLC, ≥95%
Sigma-Aldrich
Isopropyl β-D-thiogalactopyranoside solution, ReadyMade IPTG solution for Blue-white screening
Sigma-Aldrich
Arachidonic acid, from non-animal source, ≥98.5% (GC)
Supelco
2,5-Dihydroxybenzoic acid, matrix substance for MALDI-MS, ≥99.5% (HPLC), Ultra pure
Sigma-Aldrich
Dodecanoic acid, ≥99% (GC/titration)
Sigma-Aldrich
IPTG, ≥99% (TLC), ≤0.1% Dioxane
Sigma-Aldrich
Hexane, HPLC Plus, for HPLC, GC, and residue analysis, ≥95%
Sigma-Aldrich
Ethylamine solution, 2.0 M in THF
Sigma-Aldrich
Ethylamine solution, 2.0 M in methanol
Sigma-Aldrich
2,5-Dihydroxybenzoic acid, 98%
Supelco
2,5-Dihydroxybenzoic acid, matrix substance for MALDI-MS, >99.0% (HPLC)
Supelco
Hexane, suitable for HPLC, ≥95%
Sigma-Aldrich
Dodecanoic acid, 98%
Supelco
Dodecanoic acid, analytical standard
Sigma-Aldrich
Hexane, anhydrous, 95%
Sigma-Aldrich
N,O-Bis(trimethylsilyl)trifluoroacetamide, ≥99%
Sigma-Aldrich
Ethylamine solution, 66.0-72.0% in H2O
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
Arachidonylethanolamide, ≥97.0% (TLC), oil
Sigma-Aldrich
Isopropyl β-D-1-thiogalactopyranoside, ≥99% (TLC)
Sigma-Aldrich
Hexane, puriss. p.a., ACS reagent, ≥99.0% (GC)
Supelco
N,O-Bis(trimethylsilyl)trifluoroacetamide, for GC derivatization, LiChropur, ≥99.0%
Sigma-Aldrich
Arachidonic acid, >95.0% (GC)