A real-time cell-binding assay reveals dynamic features of STxB–Gb3 cointernalization and STxB-mediated cargo delivery into cancer cells

FEBS Letters

article

João Crispim Encarnação¹  Valeria Napolitano³  Giulia Opassi⁵  U. Helena Danielson⁵  Grzegorz Dubin³  Grzegorz M. Popowicz⁶  Hélène Munier-Lehmann⁸  Jos Buijs⁹  Karl Andersson¹  Hanna Björkelund¹ 

[1] Ridgeview Instruments AB;Department of Immunology, Rudbeck Laboratory, Uppsala University;Malopolska Centre of Biotechnology, Jagiellonian University;Faculty of Biochemistry, Jagiellonian University;Department of Chemistry-BMC, Uppsala University;Institute of Structural Biology;Center for Integrated Protein Science Munich at Chair of Biomolecular NMR, Department Chemie;Unité de Chimie et Biocatalyse, Institut Pasteur;Ridgeview Instruments AB, Uppsala

关键词: binding kinetics;    cancer;    cell surface receptor;    real-time cellbinding assays;    receptor internalization;    Shiga toxin;   

DOI  :  10.1002/1873-3468.13847

来源: John Wiley & Sons Ltd.

【 摘 要 】

The interaction between the Shiga toxin B-subunit (STxB) and its globotriaosylceramide receptor (Gb3) has a high potential for being exploited for targeted cancer therapy. The primary goal of this study was to evaluate the capacity of STxB to carry small molecules and proteins as cargo into cells. For this purpose, an assay was designed to provide real-time information about the StxB–Gb3 interaction as well as the dynamics and mechanism of the internalization process. The assay revealed the ability to distinguish the process of binding to the cell surface from internalization and presented the importance of receptor and STxB clustering for internalization. The overall setup demonstrated that the binding mechanism is complex, and the concept of affinity is difficult to apply. Hence, time-resolved methods, providing detailed information about the interaction of STxB with cells, are critical for the optimization of intracellular delivery.

【 授权许可】

Unknown