Home General Physiology and Biophysics 2017 General Physiology and Biophysics Vol.36, No.4, p.383–389, 2017

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Quarterly, 80 pp. per issue
Founded: 1982
ISSN  1338-4325 (online)

Published in English

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General Physiology and Biophysics Vol.36, No.4, p.383–389, 2017

Title: Cross-reacting material 197 (CRM197) affects actin cytoskeleton of endothelial cells
Author: Bilge Özerman Edis, Başak Varol, Ebru Haciosmanoğlu, Ayhan Ünlü, Muhammet Bektaş

Abstract: CRM197, cross-reacting material 197, is a mutant of diphtheria toxin (DTx). CRM197 is used in pharmacology as a carrier protein. It has been recently shown that CRM197 causes breakdown in actin filaments. In order to show intracellular localization of CRM197 and visualize cell structure via actin cytoskeleton, endothelial cells were cultured and subjected to CRM197 in vitro. To address the interaction between CRM197 and actin both experimental and theoretical studies were carried out. Colocalization of CRM197 with actin filaments was determined by immunofluorescence microscopy. Following 24-hour incubation, the loss of cell-cell contact between cells was prominent. CRM197 was shown to bind to G-actin by gel filtration chromatography, and this binding was confirmed by Western blot analysis of eluted samples obtained following chromatography. Based on crystal structure, docked model of CRM197-actin complex was generated. Molecular dynamics simulation revealed that Lys42, Cys218, Cys233 of CRM197 interacts with Gly197, Arg62 and Ser60 of G-actin, respectively. CRM197 binding to G-actin, colocalization of CRM197 with actin filament, and actin cytoskeleton rearrangement resulting in the loss of cell-cell contact show that actin comes into sight as target molecule for CRM197.

Keywords: Actin filaments — Cross-reacting material 197 — Diphtheria toxin — Endothelial cells
Published online: 13-Sep-2017
Year: 2017, Volume: 36, Issue: 4 Page From: 383, Page To: 389
doi:10.4149/gpb_2017006


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