Department of Microbiology and Immunology, Michigan Medicine, University of Michigan, Ann Arbor, Michigan, 48109 USA.
Department of Microbiology and Immunology, Michigan Medicine, University of Michigan, Ann Arbor, Michigan, 48109 USA.
J Biol Chem. 2020 Dec 4;295(49):16754-16772. doi: 10.1074/jbc.RA120.014372. Epub 2020 Sep 25.
α1-antitrypsin (AAT) regulates the activity of multiple proteases in the lungs and liver. A mutant of AAT (E342K) called ATZ forms polymers that are present at only low levels in the serum and induce intracellular protein inclusions, causing lung emphysema and liver cirrhosis. An understanding of factors that can reduce the intracellular accumulation of ATZ is of great interest. We now show that calreticulin (CRT), an endoplasmic reticulum (ER) glycoprotein chaperone, promotes the secretory trafficking of ATZ, enhancing the media:cell ratio. This effect is more pronounced for ATZ than with AAT and is only partially dependent on the glycan-binding site of CRT, which is generally relevant to substrate recruitment and folding by CRT. The CRT-related chaperone calnexin does not enhance ATZ secretory trafficking, despite the higher cellular abundance of calnexin-ATZ complexes. CRT deficiency alters the distributions of ATZ-ER chaperone complexes, increasing ATZ-BiP binding and inclusion body formation and reducing ATZ interactions with components required for ER-Golgi trafficking, coincident with reduced levels of the protein transport protein Sec31A in CRT-deficient cells. These findings indicate a novel role for CRT in promoting the secretory trafficking of a protein that forms polymers and large intracellular inclusions. Inefficient secretory trafficking of ATZ in the absence of CRT is coincident with enhanced accumulation of ER-derived ATZ inclusion bodies. Further understanding of the factors that control the secretory trafficking of ATZ and their regulation by CRT could lead to new therapies for lung and liver diseases linked to AAT deficiency.
α1-抗胰蛋白酶 (AAT) 调节肺部和肝脏中多种蛋白酶的活性。一种名为 ATZ 的 AAT 突变体 (E342K) 形成聚合物,这些聚合物在血清中的水平很低,但会诱导细胞内蛋白包涵体,导致肺气肿和肝硬化。了解可以减少 ATZ 细胞内积累的因素具有重要意义。我们现在表明,内质网 (ER) 糖蛋白伴侣钙网蛋白 (CRT) 促进 ATZ 的分泌转运,增加培养基:细胞比。与 AAT 相比,这种作用在 ATZ 中更为明显,并且仅部分依赖于 CRT 的糖结合位点,该位点通常与 CRT 对底物的募集和折叠有关。尽管 CRT-ATZ 复合物在细胞中的丰度更高,但 CRT 相关伴侣钙联蛋白 (calnexin) 并不能增强 ATZ 分泌转运。CRT 缺乏会改变 ATZ-ER 伴侣复合物的分布,增加 ATZ-BiP 结合和包涵体形成,减少 ATZ 与 ER-Golgi 转运所需成分的相互作用,同时 CRT 缺陷细胞中蛋白质转运蛋白 Sec31A 的水平降低。这些发现表明 CRT 在促进形成聚合物和大细胞内包涵体的蛋白质的分泌转运中具有新的作用。在缺乏 CRT 的情况下,ATZ 的分泌转运效率降低,与 ER 来源的 ATZ 包涵体的积累增加同时发生。进一步了解控制 ATZ 分泌转运的因素及其受 CRT 的调节可能为与 AAT 缺乏相关的肺部和肝脏疾病带来新的治疗方法。