Department of Biochemistry, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan.
J Lipid Res. 2010 Sep;51(9):2591-9. doi: 10.1194/jlr.M006049. Epub 2010 May 22.
We previously reported that the endogenous ATP-binding cassette transporter (ABC)A7 strongly associates with phagocytic function rather than biogenesis of high-density lipoprotein (HDL), being regulated by sterol-regulatory element binding protein (SREBP)2. Phagocytic activity was found enhanced by apolipoprotein (apo)A-I and apoA-II more than twice the maximum in J774 and mouse peritoneal macrophages. Therefore we investigated the molecular basis of this reaction in association with the function of ABCA7. Similar to ABCA1, ABCA7 was degraded, likely by calpain, and apoA-I and apoA-II stabilize ABCA7 against degradation. Cell surface biotinylation experiments demonstrated that endogenous ABCA7 predominantly resides on the cell surface and that the apolipoproteins increase the surface ABCA7. The increase of phagocytosis by apolipoproteins was retained in the J774 cells treated with ABCA1 siRNA and in the peritoneal macrophages from ABCA1-knockout mice, but it was abolished in the J774 cells treated with ABCA7 siRNA and in the peritoneal macrophages from ABCA7-knockout mice. Phagocytosis was decreased in the cells in the peritoneal cavity of the ABCA7-knockout mouse compared with the wild-type control. We thus concluded that extracellular helical apolipoproteins augment ABCA7-associated phagocytosis by stabilizing ABCA7. The results demonstrated direct enhancement of the host defense system by HDL components.
我们之前曾报道,内源性 ATP 结合盒转运蛋白(ABC)A7 与吞噬功能而非高密度脂蛋白(HDL)的生物发生密切相关,受固醇调节元件结合蛋白(SREBP)2 调节。载脂蛋白(apo)A-I 和 apoA-II 使 J774 和小鼠腹腔巨噬细胞的吞噬活性增强了两倍以上。因此,我们研究了与 ABCA7 功能相关的这种反应的分子基础。与 ABCA1 相似,ABCA7 被钙蛋白酶降解,而 apoA-I 和 apoA-II 稳定 ABCA7 抵抗降解。细胞表面生物素化实验表明,内源性 ABCA7 主要位于细胞表面,载脂蛋白增加了细胞表面的 ABCA7。用 ABCA1 siRNA 处理的 J774 细胞和 ABCA1 基因敲除小鼠的腹腔巨噬细胞中,载脂蛋白增加的吞噬作用得以保留,但用 ABCA7 siRNA 处理的 J774 细胞和 ABCA7 基因敲除小鼠的腹腔巨噬细胞中,这种作用被消除。与野生型对照相比,ABCA7 基因敲除小鼠腹腔细胞的吞噬作用降低。因此,我们得出结论,细胞外螺旋载脂蛋白通过稳定 ABCA7 来增强 ABCA7 相关的吞噬作用。该结果表明,HDL 成分直接增强了宿主防御系统。