Chemomechanical mapping of ligand-receptor binding kinetics on cells
Chemomechanical mapping of ligand-receptor binding kinetics on cells
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DOI：
10.1073/pnas.0702668104
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发表时间：
2007-06-05
影响因子：
11.1
通讯作者：
Van Vliet, Krystyn J.
中科院分区：
文献类型：
--
作者：
Lee, Sunyoung;Mandic, Jelena;Van Vliet, Krystyn J.
细胞表面受体和细胞外生物分子之间的结合动力学对于所有细胞内和细胞间活动至关重要。通过测量整个细胞的结合特性，可以促进受体介导的细胞功能的建模和预测，理想地指示可能影响结合受体功能的亚细胞位置或细胞骨架关联。对于配体工程和抗体中和病理过程的临床应用，这种双重需求尤其迫切。在这里，我们通过功能化力成像来绘制单个受体并确定全细胞结合动力学，这是通过扫描探针显微镜和使用生物分子共轭机械探针对完整细胞进行分子力光谱来实现的。我们量化了活体和固定人微血管内皮细胞上血管内皮生长因子受体 2 相对于单克隆抗体的数量、分布和结合/解离速率常数。这种直接受体成像的通用方法同时量化了这些受体相对于底层细胞骨架的结合动力学和不均匀分布，提供了调节受体介导行为的细胞表面动力学的时空可视化。
The binding kinetics between cell surface receptors and extracellular biomolecules is critical to all intracellular and intercellular activity. Modeling and prediction of receptor-mediated cell functions are facilitated by measurement of the binding properties on whole cells, ideally indicating the subcellular locations or cytoskeletal associations that may affect the function of bound receptors. This dual need is particularly acute vis A vis ligand engineering and clinical applications of antibodies to neutralize pathological processes. Here, we map individual receptors and determine whole-cell binding kinetics by means of functionalized force imaging, enabled by scanning probe microscopy and molecular force spectroscopy of intact cells with biomolecule-conjugated mechanical probes. We quantify the number, distribution, and association/dissociation rate constants of vascular endothelial growth factor receptor-2 with respect to a monoclonal antibody on both living and fixed human microvascular endothelial cells. This general approach to direct receptor imaging simultaneously quantifies both the binding kinetics and the nonuniform distribution of these receptors with respect to the underlying cytoskeleton, providing spatiotemporal visualization of cell surface dynamics that regulate receptor-mediated behavior.