A real‐time cell‐binding assay reveals dynamic features of STxB–Gb3 cointernalization and STxB‐mediated cargo delivery into cancer cells. Issue 15 (23rd June 2020)

• Record Type: Journal Article
• Authors: Encarnação, João Crispim; Napolitano, Valeria; Opassi, Giulia; Danielson, U. Helena; Dubin, Grzegorz; Popowicz, Grzegorz M.; Munier‐Lehmann, Hélène; Buijs, Jos; Andersson, Karl; Björkelund, Hanna
• Is Part Of: FEBS letters. Volume 594:Issue 15(2020)
• Page Start: 2406
• Page End: 2420
• Publication Date: 2020-06-23
• Subjects: binding kinetics -- cancer -- cell surface receptor -- real‐time cell‐binding assays -- receptor internalization -- Shiga toxin; Biochemistry -- Periodicals; Biophysics -- Periodicals; Molecular biology -- Periodicals; Biochimie -- Périodiques; Biochemistry; Biophysics; Molecular biology; Periodicals; 572.05
• DOI: 10.1002/1873-3468.13847
• Languages: English
• ISSNs: 0014-5793

Abstract

Abstract : 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. Abstract :