Vigliarolo Tiziana, Guida Lucrezia, Millo Enrico, Fresia Chiara, Turco Emilia, De Flora Antonio, Zocchi Elena
From the Department of Experimental Medicine, Section of Biochemistry, and
From the Department of Experimental Medicine, Section of Biochemistry, and.
J Biol Chem. 2015 May 22;290(21):13042-52. doi: 10.1074/jbc.M114.629501. Epub 2015 Apr 6.
Abscisic acid (ABA) is a plant hormone involved in the response to environmental stress. Recently, ABA has been shown to be present and active also in mammals, where it stimulates the functional activity of innate immune cells, of mesenchymal and hemopoietic stem cells, and insulin-releasing pancreatic β-cells. LANCL2, the ABA receptor in mammalian cells, is a peripheral membrane protein that localizes at the intracellular side of the plasma membrane. Here we investigated the mechanism enabling ABA transport across the plasmamembrane of human red blood cells (RBC). Both influx and efflux of [(3)H]ABA occur across intact RBC, as detected by radiometric and chromatographic methods. ABA binds specifically to Band 3 (the RBC anion transporter), as determined by labeling of RBC membranes with biotinylated ABA. Proteoliposomes reconstituted with human purified Band 3 transport [(3)H]ABA and [(35)S]sulfate, and ABA transport is sensitive to the specific Band 3 inhibitor 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid. Once inside RBC, ABA stimulates ATP release through the LANCL2-mediated activation of adenylate cyclase. As ATP released from RBC is known to exert a vasodilator response, these results suggest a role for plasma ABA in the regulation of vascular tone.
脱落酸(ABA)是一种参与植物对环境胁迫响应的植物激素。最近研究表明,ABA在哺乳动物中也存在且具有活性,它能刺激先天免疫细胞、间充质和造血干细胞以及释放胰岛素的胰腺β细胞的功能活性。LANCL2是哺乳动物细胞中的ABA受体,是一种定位于质膜胞内侧的外周膜蛋白。在此,我们研究了ABA跨人红细胞(RBC)质膜转运的机制。通过放射性测量和色谱方法检测发现,[³H]ABA的流入和流出均发生在完整的RBC上。用生物素化的ABA标记RBC膜确定,ABA特异性结合带3(RBC阴离子转运蛋白)。用人纯化的带3重构的蛋白脂质体转运[³H]ABA和[³⁵S]硫酸盐,且ABA转运对特异性带3抑制剂4,4'-二异硫氰基芪-2,2'-二磺酸敏感。一旦进入RBC,ABA通过LANCL2介导的腺苷酸环化酶激活刺激ATP释放。由于已知从RBC释放的ATP会产生血管舒张反应,这些结果表明血浆ABA在调节血管张力中起作用。