Identification of Leu276 of the S1P1 receptor and Phe263 of the S1P3 receptor in interaction with receptor specific agonists by molecular modeling, site-directed mutagenesis, and affinity studies
Identification of Leu276 of the S1P1 receptor and Phe263 of the S1P3 receptor in interaction with receptor specific agonists by molecular modeling, site-directed mutagenesis, and affinity studies
复制标题
DOI：
10.1124/mol.106.029223
复制
发表时间：
2007-03-01
影响因子：
3.6
通讯作者：
Parent, Stephen A.
中科院分区：
文献类型：
--
作者：
Deng, Qiaolin;Clemas, Joseph A.;Parent, Stephen A.
鞘氨醇-1-磷酸（S1 P）受体激动剂是新型免疫抑制剂。S1 P 1对S1 P 3的选择性与淋巴细胞隔离和最小急性毒性和心动过缓密切相关。本研究介绍了分子建模，定点诱变，和亲和力研究探索S1 P 1和S1 P 3受体之间的选择性的分子基础。基于牛视紫红质的X-射线晶体结构，建立了人S1 P1和S1 P3受体与两种非选择性激动剂或两种S1 P1选择性激动剂结合的计算模型。模型预测S1 P（1）Leu 276和S1 P（3）Phe 263对两种S1 P（1）选择性激动剂的S1 P（1）/S1 P（3）选择性有贡献。对这些残基进行定点诱变。野生型和突变型S1 P受体在中国仓鼠卵巢细胞中表达，并检查它们在体外与激动剂结合和被激动剂激活的能力。结果表明，突变对两种非选择性激动剂的活性影响很小，尽管它们对S1 P（1）-选择性激动剂有显著影响。这些研究提供了对这两种受体选择性激动剂如何与S1 P（1）和S1 P（3）受体结合的基本理解，这将有助于开发具有免疫抑制特性和改善安全性的更具选择性的S1 P（1）受体激动剂。
Sphingosine-1-phosphate (S1P) receptor agonists are novel immunosuppressive agents. The selectivity of S1P 1 against S1P 3 is strongly correlated with lymphocyte sequestration and minimum acute toxicity and bradycardia. This study describes molecular modeling, site-directed mutagenesis, and affinity studies exploring the molecular basis for selectivity between S1P 1 and S1P 3 receptors. Computational models of human S1P 1 and S1P 3 receptors bound with two nonselective agonists or two S1P 1-selective agonists were developed based on the X-ray crystal structure of bovine rhodopsin. The models predict that S1P(1) Leu276 and S1P(3) Phe263 contribute to the S1P(1)/S1P(3) selectivity of the two S1P(1)-selective agonists. These residues were subjected to site-directed mutagenesis. The wild-type and mutant S1P receptors were expressed in Chinese hamster ovary cells and examined for their abilities to bind to and be activated by agonists in vitro. The results indicate that the mutations have minimal effects on the activities of the two nonselective agonists, although they have dramatic effects on the S1P(1)-selective agonists. These studies provide a fundamental understanding of how these two receptor-selective agonists bind to the S1P(1) and S1P(3) receptors, which should aid development of more selective S1P(1) receptor agonists with immunosuppressive properties and improved safety profiles.