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A real‐time cell‐binding assay reveals dynamic features of STxB–Gb3 cointernalization and STxB‐mediated cargo delivery into cancer cells
Author(s) -
Encarnação João Crispim,
Napolitano Valeria,
Opassi Giulia,
Danielson U. Helena,
Dubin Grzegorz,
Popowicz Grzegorz M.,
MunierLehmann Hélène,
Buijs Jos,
Andersson Karl,
Björkelund Hanna
Publication year - 2020
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1002/1873-3468.13847
Subject(s) - internalization , intracellular , cancer research , cancer cell , biology , receptor , computational biology , cancer , microbiology and biotechnology , genetics
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.