Department of Microbiology and Parasitology, Faculty of Medical Science, Naresuan University, Phitsanulok, 65000, Thailand.
Graduate School of Biomedical Sciences Programme, Faculty of Allied Health Sciences, Naresuan University, Phitsanulok, 65000, Thailand.
BMC Mol Cell Biol. 2022 Jul 28;23(1):36. doi: 10.1186/s12860-022-00436-3.
The non-catalytic region of tyrosine kinase (Nck) is an adaptor protein, which is ubiquitously expressed in many types of cells. In T cells, the Nck1 isoform promotes T cell receptor signalling as well as actin polymerisation. However, the role of Nck1 in the lipid metabolism in T cells is unknown. In the present study, we investigated the effect of the Nck1 protein and Nck-CD3 interaction on lipid metabolism and on the physical and biological properties of Jurkat T cells, using a newly developed holotomographic microscope.
Holotomographic microscopy showed that Nck1-knocked-out cells had membrane blebs and were irregular in shape compared to the rounded control cells. The cell size and volume of Nck1-deficient cells were comparable to those of the control cells. Nck1-knocked-out Jurkat T cells had a greater lipid content, lipid mass/cell mass ratio, and lipid metabolite levels than the control cells. Interestingly, treatment with a small molecule, AX-024, which inhibited Nck-CD3 interaction, also caused an increase in the lipid content in wild-type Jurkat T cells, as found in Nck1-deficient cells.
Knockout of Nck1 protein and hindrance of the Nck-CD3 interaction cause the elevation of lipid content in Jurkat T cells.
酪氨酸激酶的非催化区（Nck）是一种衔接蛋白，在许多类型的细胞中广泛表达。在 T 细胞中，Nck1 同工型促进 T 细胞受体信号转导以及肌动蛋白聚合。然而，Nck1 在 T 细胞脂质代谢中的作用尚不清楚。在本研究中，我们使用新开发的全层析显微镜研究了 Nck1 蛋白和 Nck-CD3 相互作用对 Jurkat T 细胞脂质代谢以及物理和生物学特性的影响。
全层析显微镜显示，与圆形对照细胞相比，Nck1 敲除细胞具有膜泡且形状不规则。Nck1 缺陷细胞的细胞大小和体积与对照细胞相当。与对照细胞相比，Nck1 敲除的 Jurkat T 细胞具有更高的脂质含量、脂质质量/细胞质量比和脂质代谢物水平。有趣的是，用小分子 AX-024 处理抑制 Nck-CD3 相互作用，也会导致野生型 Jurkat T 细胞的脂质含量增加，这与 Nck1 缺陷细胞中的情况相似。
Nck1 蛋白的敲除和 Nck-CD3 相互作用的阻碍导致 Jurkat T 细胞中脂质含量的升高。
