Feige Matthias J, Behnke Julia, Mittag Tanja, Hendershot Linda M
From the Departments of Tumor Cell Biology and
From the Departments of Tumor Cell Biology and.
J Biol Chem. 2015 Oct 30;290(44):26821-31. doi: 10.1074/jbc.M115.689471. Epub 2015 Sep 23.
In eukaryotic cells, secretory pathway proteins must pass stringent quality control checkpoints before exiting the endoplasmic reticulum (ER). Acquisition of native structure is generally considered to be the most important prerequisite for ER exit. However, structurally detailed protein folding studies in the ER are few. Furthermore, aberrant ER quality control decisions are associated with a large and increasing number of human diseases, highlighting the need for more detailed studies on the molecular determinants that result in proteins being either secreted or retained. Here we used the clonotypic αβ chains of the T cell receptor (TCR) as a model to analyze lumenal determinants of ER quality control with a particular emphasis on how proper assembly of oligomeric proteins can be monitored in the ER. A combination of in vitro and in vivo approaches allowed us to provide a detailed model for αβTCR assembly control in the cell. We found that folding of the TCR α chain constant domain Cα is dependent on αβ heterodimerization. Furthermore, our data show that some variable regions associated with either chain can remain incompletely folded until chain pairing occurs. Together, these data argue for template-assisted folding at more than one point in the TCR α/β assembly process, which allows specific recognition of unassembled clonotypic chains by the ER chaperone machinery and, therefore, reliable quality control of this important immune receptor. Additionally, it highlights an unreported possible limitation in the α and β chain combinations that comprise the T cell repertoire.
在真核细胞中,分泌途径蛋白在离开内质网(ER)之前必须通过严格的质量控制检查点。获得天然结构通常被认为是内质网输出的最重要前提条件。然而,在内质网中进行的结构详细的蛋白质折叠研究却很少。此外,异常的内质网质量控制决策与大量且不断增加的人类疾病相关,这凸显了对导致蛋白质分泌或保留的分子决定因素进行更详细研究的必要性。在这里,我们使用T细胞受体(TCR)的克隆型αβ链作为模型来分析内质网质量控制的腔内决定因素,特别强调如何在内质网中监测寡聚蛋白的正确组装。体外和体内方法的结合使我们能够为细胞中的αβTCR组装控制提供一个详细模型。我们发现TCRα链恒定区Cα的折叠依赖于αβ异二聚体化。此外,我们的数据表明,与任一链相关的一些可变区在链配对发生之前可能仍未完全折叠。总之,这些数据支持在TCRα/β组装过程中的多个点进行模板辅助折叠,这使得内质网伴侣机制能够特异性识别未组装的克隆型链,从而对这一重要免疫受体进行可靠的质量控制。此外,它还突出了构成T细胞库的α链和β链组合中一个未报道的可能限制。