Chemically engineering ligand selectivity at the free fatty acid receptor 2 based on pharmacological variation between species orthologs.
Chemically engineering ligand selectivity at the free fatty acid receptor 2 based on pharmacological variation between species orthologs.
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DOI：
10.1096/fj.12-213314
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发表时间：
2012-12
期刊：
影响因子：
--
通讯作者：
Milligan G
中科院分区：
文献类型：
--
作者：
Hudson BD;Christiansen E;Tikhonova IG;Grundmann M;Kostenis E;Adams DR;Ulven T;Milligan G
当难以在相关G蛋白偶联受体（GPCR）家族内开发选择性配体时，仅由合成配体（RASSL）激活的化学工程受体是探测受体功能的有用替代方案。在目前的工作中，我们探讨了是否RASSL的游离脂肪酸受体2（FFA 2）可以开发的基础上物种直系同源物之间的药理学差异。为此，牛FFA 2进行了表征，揭示了与人FFA 2相比不同的配体选择性。同源性建模和突变分析表明，C4.57G的人FFA 2中的单个突变导致具有与牛受体相似的配体选择性的人FFA 2受体。通过在已知的正构配体相互作用位点H6.55Q添加第二突变，利用这一点来产生人FFA 2-RASSL。所得的FFA 2-RASSL显示出对内源性配体的>100倍的活性损失，同时响应于不同的配体山梨酸，其中cAMP抑制的PEC 50值为5.83 ± 0.11; Ca 2+动员，4.63 ± 0.05; ERK磷酸化，5.61 ± 0.06;以及动态质量再分布，5.35 ± 0.06。这种FFA 2-RASSL将在未来对这种受体的研究中有用，并表明利用物种直系同源物之间的药理学差异是产生新型化学工程GPCR的有力方法。哈德逊，B。D、Christiansen，E.，吉洪诺娃岛G.，Grundmann，M.，Kostenis，E.，亚当斯，D. R.，Ulven，T.，米利根湾，澳-地基于物种直系同源物之间的药理学差异，化学工程化游离脂肪酸受体2的配体选择性。
When it is difficult to develop selective ligands within a family of related G-protein-coupled receptors (GPCRs), chemically engineered receptors activated solely by synthetic ligands (RASSLs) are useful alternatives for probing receptor function. In the present work, we explored whether a RASSL of the free fatty acid receptor 2 (FFA2) could be developed on the basis of pharmacological variation between species orthologs. For this, bovine FFA2 was characterized, revealing distinct ligand selectivity compared with human FFA2. Homology modeling and mutational analysis demonstrated a single mutation in human FFA2 of C4.57G resulted in a human FFA2 receptor with ligand selectivity similar to the bovine receptor. This was exploited to generate human FFA2-RASSL by the addition of a second mutation at a known orthosteric ligand interaction site, H6.55Q. The resulting FFA2-RASSL displayed a >100-fold loss of activity to endogenous ligands, while responding to the distinct ligand sorbic acid with pEC50 values for inhibition of cAMP, 5.83 ± 0.11; Ca2+ mobilization, 4.63 ± 0.05; ERK phosphorylation, 5.61 ± 0.06; and dynamic mass redistribution, 5.35 ± 0.06. This FFA2-RASSL will be useful in future studies on this receptor and demonstrates that exploitation of pharmacological variation between species orthologs is a powerful method to generate novel chemically engineered GPCRs.—Hudson, B. D., Christiansen, E., Tikhonova, I. G., Grundmann, M., Kostenis, E., Adams, D. R., Ulven, T., Milligan, G. Chemically engineering ligand selectivity at the free fatty acid receptor 2 based on pharmacological variation between species orthologs.
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DOI：
10.1073/pnas.0700293104
发表时间：
2007-03-20
影响因子：
11.1
作者：
Armbruster, Blaine N.;Li, Xiang;Roth, Bryan L.
通讯作者：
Roth, Bryan L.
DOI：
10.1111/j.1463-1326.2009.01114.x
发表时间：
2009-11
期刊：
Diabetes, obesity & metabolism
影响因子：
--
作者：
Kebede MA;Alquier T;Latour MG;Poitout V
通讯作者：
Poitout V
影响因子：
14.8
作者：
Schroeder, Ralf;Schmidt, Johannes;Kostenis, Evi
通讯作者：
Kostenis, Evi
影响因子：
64.8
作者：
He, WH;Miao, FJP;Ling, L
通讯作者：
Ling, L
DOI：
10.1016/b978-0-12-385952-5.00011-7
发表时间：
2011-01-01
期刊：
PHARMACOLOGY OF G PROTEIN COUPLED RECEPTORS
影响因子：
--
作者：
Congreve, Miles;Langmead, Christopher;Marshall, Fiona H.
通讯作者：
Marshall, Fiona H.