Muntasir Habib Abul, Hossain Murad, Bhuiyan Mohiuddin Ahmed, Komiyama Tadazumi, Nakamura Takashi, Ozaki Masanobu, Nagatomo Takafumi
Department of Pharmacology, Faculty of Pharmaceutical Sciences, Niigata University of Pharmacy and Applied Life Sciences, 265-1 Higashijima, Akihaku, Niigata 956-8603, Japan.
J Pharmacol Sci. 2007 Jul;104(3):274-7. doi: 10.1254/jphs.sc0060241. Epub 2007 Jul 3.
Based on radio-ligand binding and molecular modeling studies, sarpogrelate shows a moderate selectivity for 5-HT(2B) versus 5-HT(2A) receptors. To confirm the modeling data of sarpogrelate to 5-HT(2B) receptors predicting interaction of sarpogrelate towards Asp135 in helix 3 of 5-HT(2B) receptors, we constructed and characterized the mutation of this residue by site-directed mutagenesis. The Asp135Ala mutant did not exhibit any affinity for [(3)H]rauwolscine. Therefore, it was not possible to find sarpogrelate affinity to the mutant using [(3)H]rauwolscine. The mutation also abolished agonist-stimulated inositol phosphates formation. These results provide evidence that Asp135 is important for the interaction between 5-HT(2B) receptors and sarpogrelate.
基于放射性配体结合和分子模拟研究，相对于5-HT(2A)受体，沙格雷酯对5-HT(2B)受体表现出适度的选择性。为了证实沙格雷酯与5-HT(2B)受体的模拟数据，该数据预测了沙格雷酯与5-HT(2B)受体第3螺旋中Asp135的相互作用，我们通过定点诱变构建并表征了该残基的突变。Asp135Ala突变体对[(3)H]萝芙木碱没有任何亲和力。因此，使用[(3)H]萝芙木碱无法检测到沙格雷酯对该突变体的亲和力。该突变还消除了激动剂刺激的肌醇磷酸生成。这些结果证明，Asp135对于5-HT(2B)受体与沙格雷酯之间的相互作用很重要。
