Journal Article Epublish
PubMed: 40443899 DOI: 10.1021/jacsau.5c00193
Characterizing the Membrane Assembly of ASGPR Related to Mediated Endocytosis Using TriGalNAc-Probe-Based Super-Resolution Imaging.
JACS Au 5(5):2246-2256
Authors: Junling Chen, Jiaqi Wang, Binglin Sui, Pengwei Jiang, Xumin Wang, Hongda Wang, Feng Liang
Affiliations
- School of Chemistry & Chemical Engineering, Wuhan University of Science and Technology, 947 Heping Street, Wuhan, Hubei 430081, China.
- College of Chemical Engineering and Environmental Chemistry, Weifang University, Weifang 261061, China.
- State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Research Center of Biomembranomics, 5625 Renmin Street, Changchun, Jilin 130022, China.
Abstract
The asialoglycoprotein receptor (ASGPR) is a promising therapeutic target for drug delivery systems in hepatocellular carcinoma (HCC), exhibiting high affinity for specific carbohydrate residues and overexpression on malignant hepatic cells. However, their functional mechanisms remain poorly resolved at the single molecule level, hindering the rational optimization of ASGPR-targeted drug delivery systems. Here, we developed a trivalent N-acetylgalactosamine (TriGalNAc)-functionalized ligand probe leveraging high affinity to enable the nanoscale visualization of ASGPR organization and trafficking via super-resolution imaging. Fixed cell imaging revealed pronounced clustering patterns of the ASGPR on HCC membranes. In live cell experiments, we observed the distribution changes of residual ASGPR and returned ASGPR on the membrane during endocytosis, identifying protein clusters as key functional platforms for mediated ligand uptake. Additionally, comparisons with ligand probe binding under varying cell states confirmed that ASGPR aggregation degree correlates with its ligand-binding capacity. Strikingly, disruption of membrane carbohydrate cross-linking dispersed ASGPR clusters and attenuated ligand binding. These findings resolve the nanoscale assembly of ASGPR in HCC and unveil clustering-dependent ligand-binding regulation, advancing a fundamental understanding of ASGPR biology while providing new insights to refine receptor-targeted therapeutics.
MeSH
Cells, Pharmaceutical Preparations, Cluster Analysis, Drug Delivery Systems, Therapeutics, Carbohydrates, Ligands, Membranes, Endocytosis