Perlmutter D H, Daniels J D, Auerbach H S, De Schryver-Kecskemeti K, Winter H S, Alpers D H
Department of Pediatrics, Washington University School of Medicine, St. Louis, Missouri.
J Biol Chem. 1989 Jun 5;264(16):9485-90.
alpha 1-Antitrypsin (alpha 1-AT) is considered a typical plasma protein and a prototype of the serine proteinase inhibitor (serpin) family. It is synthesized in hepatocytes and, to a lesser extent, in macrophages. In this study we show that the alpha 1-AT gene is also expressed in human intestine and in a human colonic epithelial tumor cell line, Caco2. A single 1.6-kilobase alpha 1-AT-specific mRNA is present in jejunum and in Caco2 cells. It is identical in apparent size to that present in human hepatoma HepG2 cells but slightly smaller than that present in human macrophages, cells in which an alternative upstream transcriptional start site is used. Synthesis and secretion of alpha 1-AT in Caco2 cells is similar to that in HepG2 cells. It is synthesized as an approximately 52-kDa precursor polypeptide, converted to its mature, fully glycosylated 55-kDa form intracellularly, and the native protein is secreted with a half-time of 37 min. Functionally active alpha 1-AT is secreted into the basolateral and apical (luminal) fluid in pulse-chase labeling experiments of Caco2 cells cultured in polarized orientation on collagen-coated nitrocellulose membranes. Expression of alpha 1-AT in Caco2 enterocytes is not affected by soluble factors that regulate expression of alpha 1-AT in macrophages and hepatocytes. However, expression of alpha 1-AT increases markedly in Caco2 cells as they differentiate into enteric villous-type cells.
α1抗胰蛋白酶(α1-AT)被认为是一种典型的血浆蛋白,也是丝氨酸蛋白酶抑制剂(丝氨酸蛋白酶抑制剂)家族的原型。它在肝细胞中合成,在巨噬细胞中的合成量较少。在本研究中,我们发现α1-AT基因也在人肠道和人结肠上皮肿瘤细胞系Caco2中表达。空肠和Caco2细胞中存在单一的1.6千碱基α1-AT特异性mRNA。其表观大小与人肝癌HepG2细胞中的相同,但略小于人巨噬细胞中的,在人巨噬细胞中使用了另一个上游转录起始位点。Caco2细胞中α1-AT的合成和分泌与HepG2细胞中的相似。它作为一种约52 kDa的前体多肽合成,在细胞内转化为其成熟的、完全糖基化的55 kDa形式,天然蛋白以37分钟的半衰期分泌。在胶原包被的硝酸纤维素膜上以极化方向培养的Caco2细胞的脉冲追踪标记实验中,功能活性α1-AT分泌到基底外侧和顶端(管腔)液中。Caco2肠上皮细胞中α1-AT的表达不受调节巨噬细胞和肝细胞中α1-AT表达的可溶性因子的影响。然而,随着Caco2细胞分化为肠绒毛型细胞,α1-AT的表达显著增加。