Effects of flurbiprofen on the functional regulation of serotonin transporter and its misfolded mutant
Effects of flurbiprofen on the functional regulation of serotonin transporter and its misfolded mutant
复制标题
氟比洛芬对血清素转运蛋白及其错误折叠突变体功能调节的影响
DOI：
10.1016/j.jphs.2021.11.006
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发表时间：
2022
影响因子：
3.5
通讯作者：
Sakai Norio
中科院分区：
文献类型：
--
作者：
Hirakawa Haruki;Taguchi Kei;Murakawa Seiya;Asano Masaya;Noguchi Soma;Kikkawa Satoshi;Harada Kana;Adachi Naoko;Ueyama Takehiko;Hide Izumi;Tanaka Shigeru;Sakai Norio
氟比洛芬是一种非甾体抗炎药，据报道具有化学伴侣活性。在此，我们研究了氟比洛芬通过膜运输调节5-羟色胺转运体（SERT）功能的作用。我们使用瞬时表达野生型（WT）SERT或SERT C-末端缺失突变体（SERTΔCT）（一种错误折叠蛋白）的COS-7细胞。氟比洛芬处理降低了非糖基化SERT的表达，增强了非糖基化SERT的表达。此外，我们观察到SERT表达细胞中5-羟色胺摄取增加。这些结果表明，氟比洛芬调节SERT功能，促进膜运输。在表达SERTΔ CT的细胞中，氟比洛芬降低了SERTΔCT的蛋白表达和摄取活性。此外，氟比洛芬抑制了SERTΔCT聚集体的形成。使用氟比洛芬对映体的研究表明，氟比洛芬对SERT的这些作用不是通过环氧合酶抑制介导的。评价内质网应激标志物GRP 78/BiP的水平以阐明氟比洛芬是否可以改善SERTΔ CT诱导的内质网应激。有趣的是，氟比洛芬诱导GRP 78/BiP表达仅在ER应激条件下，而不是在稳态条件下。在HRD 1 E3泛素连接酶敲低细胞中，氟比洛芬影响ER相关降解系统。总的来说，研究结果表明，氟比洛芬可能作为一个诱导剂的分子伴侣，除了作为一个化学伴侣。
Flurbiprofen, a nonsteroidal anti-inflammatory drug, reportedly exhibits chemical chaperone activity. Herein, we investigated the role of flurbiprofen in regulating serotonin transporter (SERT) function via membrane trafficking. We used COS-7 cells transiently expressing wild-type (WT) SERT or a C-terminus-deleted mutant of SERT (SERTΔCT), a misfolded protein. Flurbiprofen treatment reduced the expression of immaturely glycosylated SERT and enhanced the expression of maturely glycosylated SERT. In addition, we observed increased serotonin uptake in SERT-expressing cells. These results suggest that flurbiprofen modulates SERT function by promoting membrane trafficking. In SERTΔCT-expressing cells, flurbiprofen reduced the protein expression and uptake activity of SERTΔCT. Furthermore, flurbiprofen inhibited the formation of SERTΔCT aggregates. Studies using flurbiprofen enantiomers suggested that these effects of flurbiprofen on SERT were not mediated via cyclooxygenase inhibition. The levels of GRP78/BiP, an endoplasmic reticulum (ER) stress marker, were assessed to elucidate whether flurbiprofen can ameliorate SERTΔCT-induced ER stress. Interestingly, flurbiprofen induced GRP78/BiP expression only under ER stress conditions and not under steady-state conditions. In HRD1 E3 ubiquitin ligase knockdown cells, flurbiprofen affected the ER-associated degradation system. Collectively, the findings suggest that flurbiprofen may function as an inducer of molecular chaperones, in addition to functioning as a chemical chaperone.