<br>使用单分子力光谱法对单个细胞表面的磷脂酰丝氨酸分子进行定量,Analytical Chemistry

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使用单分子力光谱法对单个细胞表面的磷脂酰丝氨酸分子进行定量
Analytical Chemistry ( IF 6.7 ) Pub Date : 2024-01-03 , DOI: 10.1021/acs.analchem.3c03517
Zhirong Li ₁ , Lulu Zhang ₁ , Zhanzhong Wang ₁ , Xiongli Kang ₁ , Huiying Jin ₁ , Wenjie Zhao ₁ , Jun Zhang ₁ , Haiquan Su ₁

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鉴定细胞凋亡过程中细胞膜上发生的磷脂酰丝氨酸 (PS) 差异至关重要。然而，在单细胞水平上实时监测PS分子的数量目前仍然是一项具有挑战性的任务。在这里，我们通过利用二吡啶胺锌 (II) 复合物 (ZnDPA) 与 PS 所表现出的特异性结合和可逆相互作用来证明这一目标。随后将硫辛酸功能化的 ZnDPA (LP-ZnDPA) 固定在原子力显微镜悬臂的表面上，形成力探针 ALP-ZnDPA，从而实现 PS 特异性的动态成像和检测模式。通过利用这项技术，我们不仅可以创建亚微米分辨率的 PS 表达水平热图，还可以量化单个细胞表面存在的分子数量，检测限为 1.86 × 10 ^{4 个}分子。通过分析不同癌细胞系和紫杉醇诱导细胞凋亡的各个阶段的 PS 表达水平，证明了该方法的可行性。这项研究首次应用力探针来量化单个细胞表面的 PS 分子，在分子水平上深入了解细胞凋亡过程中 PS 含量的动态变化，并为当前的检测方法引入了新的维度。

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Quantification of Phosphatidylserine Molecules on the Surface of Individual Cells Using Single-Molecule Force Spectroscopy

Identification of the phosphatidylserine (PS) discrepancies occurring on the cellular membrane during apoptotic processes is of the utmost importance. However, monitoring the quantity of PS molecules in real-time at a single-cell level currently remains a challenging task. Here, we demonstrate this objective by leveraging the specific binding and reversible interaction exhibited by the zinc(II) dipyridinamine complex (ZnDPA) with PS. Lipoic acid-functionalized ZnDPA (LP-ZnDPA) was subsequently immobilized onto the surface of an atomic force microscopy cantilever to form a force probe, ALP–ZnDPA, enabling a PS-specific dynamic imaging and detection mode. By utilizing this technique, we can not only create a heat map of the expression level of PS with submicron resolution but also quantify the number of molecules present on a single cell’s surface with a detection limit of 1.86 × 10⁴ molecules. The feasibility of the proposed method is demonstrated through the analysis of PS expression levels in different cancer cell lines and at various stages of paclitaxel-induced apoptosis. This study represents the first application of a force probe to quantify PS molecules on the surface of individual cells, providing insight into dynamic changes in PS content during apoptosis at the molecular level and introducing a novel dimension to current detection methodologies.

更新日期：2024-01-03

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