Biotechnology Institute Thurgau (BITg), University of Konstanz, CH-8280 Kreuzlingen, Switzerland.
Graduate School for Cellular and Biomedical Sciences, University of Bern, CH-3012 Bern, Switzerland.
Cells. 2022 Apr 25;11(9):1444. doi: 10.3390/cells11091444.
The chemokine receptor CCR7, together with its ligands, is responsible for the migration and positioning of adaptive immune cells, and hence critical for launching adaptive immune responses. CCR7 is also induced on certain cancer cells and contributes to metastasis formation. Thus, CCR7 expression and signalling must be tightly regulated for proper function. CCR7, like many other members of the G-protein coupled receptor superfamily, can form homodimers and oligomers. Notably, danger signals associated with pathogen encounter promote oligomerisation of CCR7 and is considered as one layer of regulating its function. Here, we assessed the dimerisation of human CCR7 and several single point mutations using split-luciferase complementation assays. We demonstrate that dimerisation-defective CCR7 mutants can be transported to the cell surface and elicit normal chemokine-driven G-protein activation. By contrast, we discovered that CCR7 mutants whose expression are shifted towards monomers significantly augment their capacities to bind and internalise fluorescently labelled CCL19. Modeling of the receptor suggests that dimerisation-defective CCR7 mutants render the extracellular loops more flexible and less structured, such that the chemokine recognition site located in the binding pocket might become more accessible to its ligand. Overall, we provide new insights into how the dimerisation state of CCR7 affects CCL19 binding and receptor trafficking.
趋化因子受体 CCR7 与其配体一起负责适应性免疫细胞的迁移和定位,因此对启动适应性免疫反应至关重要。CCR7 也在某些癌细胞上诱导,并有助于转移形成。因此,CCR7 的表达和信号必须受到严格调控,以确保其正常功能。CCR7 像 G 蛋白偶联受体超家族的许多其他成员一样,可以形成同源二聚体和寡聚体。值得注意的是,与病原体接触相关的危险信号促进 CCR7 的寡聚化,被认为是调节其功能的一层。在这里,我们使用荧光素酶互补测定法评估了人 CCR7 及其几种单点突变体的二聚化。我们证明,二聚化缺陷的 CCR7 突变体可以被转运到细胞膜表面,并引发正常的趋化因子驱动的 G 蛋白激活。相比之下,我们发现,其表达偏向单体的 CCR7 突变体显著增强了它们结合和内化荧光标记的 CCL19 的能力。受体的建模表明,二聚化缺陷的 CCR7 突变体使细胞外环更加灵活和不稳定,使得位于结合口袋中的趋化因子识别位点可能更容易被其配体结合。总的来说,我们提供了新的见解,了解 CCR7 的二聚化状态如何影响 CCL19 的结合和受体转运。