Substrate Specificity and Stereoselectivity of Rat Brain Microsomal Anandamide Amidohydrolase

Lang, Wensheng; Qin, Ce; Lin, Sonyuan; Khanolkar, Atmaram D.; Goutopoulos, Andreas; Fan, Pusheng; Abouzid, Khaled A. M.; Meng, Ziqiang; Biegel, Diane; Makriyannis, Alexandros

doi:10.1021/jm990139o

Cited by 39 publications

(55 citation statements)

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“…2). It is interesting to note that the enantioselectivity for anandamide transport displayed by 20 and 21 and 33 and 34 is congruent to that demonstrated for anandamide amidohydrolase, but opposite to that for the CB1 receptor (20,25).…”

mentioning

confidence: 71%

Structural determinants for recognition and translocation by the anandamide transporter

Piomelli

Beltramo

Glasnapp

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

251

205

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The biological actions of anandamide (arachidonylethanolamide), an endogenous cannabinoid lipid, are terminated by a two-step inactivation process consisting of carrier-mediated uptake and intracellular hydrolysis. Anandamide uptake in neurons and astrocytes is mediated by a high-affinity, Na ؉ -independent transporter that is selectively inhibited by N-(4-hydroxyphenyl)-arachidonamide (AM404). In the present study, we examined the structural determinants governing recognition and translocation of substrates by the anandamide transporter constitutively expressed in a human astrocytoma cell line. Competition experiments with a select group of analogs suggest that substrate recognition by the transporter is favored by a polar nonionizable head group of defined stereochemical configuration containing a hydroxyl moiety at its distal end. The secondary carboxamide group interacts favorably with the transporter, but may be replaced with either a tertiary amide or an ester, suggesting that it may serve as hydrogen acceptor. Thus, 2-arachidonylglycerol, a putative endogenous cannabinoid ester, also may serve as a substrate for the transporter. Substrate recognition requires the presence of at least one cis double bond situated at the middle of the fatty acid carbon chain, indicating a preference for ligands whose hydrophobic tail can adopt a bent U-shaped conformation. On the other hand, uptake experiments with radioactively labeled substrates show that no fewer than four cis nonconjugated double bonds are required for optimal translocation across the cell membrane, suggesting that substrates are transported in a folded hairpin conformation. These results outline the general structural requisites for anandamide transport and may assist in the development of selective inhibitors with potential clinical applications.

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mentioning

confidence: 71%

Structural determinants for recognition and translocation by the anandamide transporter

Piomelli

Beltramo

Glasnapp

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

251

205

View full text Add to dashboard Cite

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“…Moreover, endogenous amide hydrolases do not readily cleave fatty acyl tertiary amides, i.e. those with two substitutions on the amide nitrogen (18). Thus, attaching the cage to the amide nitrogen of VNA should generate a caged molecule that is both biologically inactive and resistant to enzymatic degradation.…”

Section: Resultsmentioning

confidence: 99%

Caged Vanilloid Ligands for Activation of TRPV1 Receptors by 1- and 2-Photon Excitation

et al. 2006

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Nociceptive neurons in the peripheral nervous system detect noxious stimuli and report the information to the central nervous system. Most nociceptive neurons express the vanilloid receptor, TRPV1, a non-selective cation channel gated by vanilloid ligands such as capsaicin, the pungent essence of chili peppers. Here, we report the synthesis and biological application of two caged vanilloids-biologically inert precursors that, when photolyzed, release bioactive vanilloid ligands. The two caged vanilloids, Nb-VNA and Nv-VNA, are photoreleased with quantum efficiency of 0.13 and 0.041, respectively. Under flash photolysis conditions, photorelease of Nb-VNA and Nv-VNA is 95% complete in ∼40 μs and ∼125 μs, respectively. Through 1-photon excitation with ultraviolet light (360 nm), or 2-photon excitation with red light (720 nm), the caged vanilloids can be photoreleased in situ to activate TRPV1 receptors on nociceptive neurons. The consequent increase in intracellular free Ca 2+ concentration ([Ca 2+ ] i ) can be visualized by laser-scanning confocal imaging of neurons loaded with the fluorescent Ca 2+ indicator, fluo-3. Stimulation results from TRPV1 receptor activation, because the response is blocked by capsazepine, a selective TRPV1 antagonist. In Ca 2+ -free extracellular medium, photoreleased vanilloid can still elevate [Ca 2+ ] i , which suggests that TRPV1 receptors also reside on endomembranes in neurons and can mediate Ca 2+ release from intracellular stores. Notably, whole-cell voltage clamp measurements showed that flash photorelease of vanilloid can activate TRPV1 channels in < 4 msec at 22°C. In combination with 1-or 2-photon excitation, caged vanilloids are a powerful tool for probing morphologically distinct structures of nociceptive sensory neurons with high spatial and temporal precision. Keywords photolysis; photorelease; uncaging; molecular probes In the peripheral nervous system, nociceptive sensory neurons, or nociceptors, detect noxious stimuli and convert these stimuli into action potentials that are transmitted to the central nervous system (1). The vast majority of nociceptive neurons express the vanilloid receptor, TRPV1, 2 which is a ligand-gated non-selective cation channel with high permeability to

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“…This was somewhat surprising, given the broad range of acyl amides and acyl esters that can be hydrolyzed by FAAH (3). On the other hand, previous studies demonstrated that substitutions at the ␣-position of the acyl chain of primary amide or anandamide analogue compounds rendered the compound resistant to hydrolysis by FAAH (39,40). The incorporation of a bulky phenoxy-group near the amide moiety may serve a similar structural hindrance to the enzyme's active site and restrict hydrolysis.…”

Section: Discussionmentioning

confidence: 99%

Synthesis of Phenoxyacyl-Ethanolamides and Their Effects on Fatty Acid Amide Hydrolase Activity

Faure

Nagarajan

Hwang

et al. 2014

Journal of Biological Chemistry

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Background: Fatty acid amide hydrolase (FAAH) belongs to the family of amidase signature proteins and is involved in N-acylethanolamine (NAE) metabolism. Results: New synthetic phenoxyacyl-ethanolamide compounds increased the amidohydrolase activity of FAAH. Conclusion: Phenoxyacyl-ethanolamide compounds increased the activity of the FAAH enzyme in plants and animals. Significance: New properties of FAAH proteins were revealed with these phenoxyacyl-ethanolamide compounds, and the potential for their applications in vivo was demonstrated.

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Substrate Specificity and Stereoselectivity of Rat Brain Microsomal Anandamide Amidohydrolase

Abstract: In Table 2, the IC 50 's of compounds 3 and 4 should be switched: compound 3, >50 a ; compound 4, 13.3 a .

Cited by 39 publications

References 0 publications

Structural determinants for recognition and translocation by the anandamide transporter

Structural determinants for recognition and translocation by the anandamide transporter

Caged Vanilloid Ligands for Activation of TRPV1 Receptors by 1- and 2-Photon Excitation

Synthesis of Phenoxyacyl-Ethanolamides and Their Effects on Fatty Acid Amide Hydrolase Activity

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