Structural determinants for recognition and translocation by the anandamide transporter.
Structural determinants for recognition and translocation by the anandamide transporter.
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anandamide 转运蛋白识别和易位的结构决定因素。
DOI：
10.1073/pnas.96.10.5802
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发表时间：
1999
影响因子：
11.1
通讯作者：
Makriyannis,A
中科院分区：
文献类型：
--
作者：
Piomelli,D;Beltramo,M;Glasnapp,S;Lin,SY;Goutopoulos,A;Xie,XQ;Makriyannis,A
内源性大麻素脂质花生四烯酸乙醇酰胺（arachidonyethanolamide）的生物学作用由载体介导的摄取和细胞内水解组成的两步失活过程终止。神经元和星形胶质细胞对花生四烯酸的摄取是由一种高亲和力的Na+非依赖性转运蛋白介导的，该转运蛋白可被N-（4-羟基苯基）-花生四烯酸酰胺（AM404）选择性抑制。在本研究中，我们研究了由花生四烯酸转运蛋白在人星形细胞瘤细胞系中组成型表达的底物识别和转运的结构决定因素。与一组选择的类似物的竞争实验表明，底物识别的转运蛋白是由一个极性的不可电离的头部基团的定义的立体化学配置，在其远端含有羟基部分的青睐。二级羧酰胺基团与转运蛋白有利地相互作用，但可以被叔酰胺或酯取代，这表明它可以作为氢受体。因此，2-花生四烯酸甘油，一种假定的内源性大麻素酯，也可以作为转运蛋白的底物。底物识别需要至少一个顺式双键的存在下，位于中间的脂肪酸碳链，表明优选的配体，其疏水尾部可以采取弯曲的U形构象。另一方面，放射性标记的底物的摄取实验表明，不少于四个顺式非共轭双键需要跨细胞膜的最佳易位，这表明底物在折叠的发夹构象运输。这些结果概述了anandamide运输的一般结构，并可能有助于开发具有潜在临床应用的选择性抑制剂。
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 byN-(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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影响因子：
5
作者：
Calignano, A;LaRana, G;Piomelli, D
通讯作者：
Piomelli, D
DOI：
10.1073/pnas.95.15.8625
发表时间：
1998-07-21
影响因子：
11.1
作者：
Hirsch, D;Stahl, A;Lodish, HF
通讯作者：
Lodish, HF
猪脑酶水解和合成 Anandamide 的部分纯化和表征 (*)
DOI：
10.1074/jbc.270.40.23823
发表时间：
1995
期刊：
The Journal of Biological Chemistry
影响因子：
--
作者：
N. Ueda;Y. Kurahashi;Shozo Yamamoto;T. Tokunaga
通讯作者：
T. Tokunaga
反异脂肪酸 1- 和 2- 单甘油酯的合成和表征。
影响因子：
6.5
作者：
B. Serdarevich;K. Carroll
通讯作者：
K. Carroll
1-或3-酰基-sn-甘油合成的改进方法。
影响因子：
6.5
作者：
Kodali,DR
通讯作者：
Kodali,DR