Wei Yushuang, Li Xiangyang, Gong Yao, Li Yue-Xuan, Guan Jibin, Yuan Bing, Chen Yue, Pang Hong-Bo
Department of Pharmaceutics, University of Minnesota, Minneapolis, MN 55455, USA.
Songshan Lake Materials Laboratory, Dongguan, Guangdong, 523808, China.
Nano Today. 2025 Dec;65. doi: 10.1016/j.nantod.2025.102865. Epub 2025 Aug 9.
Nanomaterials often need to interact with proteins on the plasma membrane to get cross and access their intracellular targets. Therefore, to fully understand the cell entry mechanism, it is of vital importance to gain a comprehensive insight into the proteome at the interface when nanomaterials encounter the cells. Here, we reported a peroxidase-based proximity labeling method to survey the proteome at the nanoparticle (NP)-cell interface. Horseradish peroxidase (HRP) was conjugated to a variety of NPs and other ligand types while still being able to biotinylate the proteins surrounding NP (or ligand)-receptor complexes. Using two NP-based tracers for macropinocytosis (MP), which is highly relevant to NP internalization, we performed a proteomic survey and revealed the interface proteome difference between traditional and receptor-dependent MP. Moreover, our survey found that E-cadherin (CDH1), while not serving as the primary receptor, is present at the NP-cell interface and is functionally important for the cellular uptake of a wide variety of NPs. Overall, by integrating nanotechnology with proximity labeling, our study provides an approach to map the proteome of NP-cell interface for investigating the molecular mechanism of NP and macromolecule internalization into cells.
纳米材料通常需要与质膜上的蛋白质相互作用，以穿过并进入其细胞内靶点。因此，为了全面了解细胞进入机制，当纳米材料与细胞相遇时，深入洞察界面处的蛋白质组至关重要。在此，我们报道了一种基于过氧化物酶的邻近标记方法，用于检测纳米颗粒（NP）-细胞界面处的蛋白质组。辣根过氧化物酶（HRP）与多种纳米颗粒及其他配体类型偶联，同时仍能够对纳米颗粒（或配体）-受体复合物周围的蛋白质进行生物素化。使用两种基于纳米颗粒的巨胞饮作用（MP）示踪剂（巨胞饮作用与纳米颗粒内化高度相关），我们进行了蛋白质组学检测，揭示了传统巨胞饮作用和受体依赖性巨胞饮作用之间的界面蛋白质组差异。此外，我们的检测发现，E-钙黏蛋白（CDH1）虽然不是主要受体，但存在于纳米颗粒-细胞界面，并且对多种纳米颗粒的细胞摄取具有重要功能。总体而言，通过将纳米技术与邻近标记相结合，我们的研究提供了一种绘制纳米颗粒-细胞界面蛋白质组图谱的方法，用于研究纳米颗粒和大分子进入细胞内吞作用的分子机制。
