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Pitstop‐2 and its novel derivative RVD‐127 disrupt global cell dynamics and nuclear pores integrity by direct interaction with small GTPases

Clathrin‐mediated endocytosis (CME) is an essential cell physiological process of broad biomedical relevance. Since the recent introduction of Pitstop‐2 as a potent CME inhibitor, we and others have reported on substantial clathrin‐independent inhibitory effects. Herein, we developed and experimenta...

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Autores principales: Liashkovich, Ivan, Stefanello, Sílvio Terra, Vidyadharan, Reshma, Haufe, Günter, Erofeev, Alexander, Gorelkin, Peter V., Kolmogorov, Vasilii, Mizdal, Caren Rigon, Dulebo, Alexander, Bulk, Etmar, Kouzel, Ian U., Shahin, Victor
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley & Sons, Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10354767/
https://www.ncbi.nlm.nih.gov/pubmed/37476059
http://dx.doi.org/10.1002/btm2.10425
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author Liashkovich, Ivan
Stefanello, Sílvio Terra
Vidyadharan, Reshma
Haufe, Günter
Erofeev, Alexander
Gorelkin, Peter V.
Kolmogorov, Vasilii
Mizdal, Caren Rigon
Dulebo, Alexander
Bulk, Etmar
Kouzel, Ian U.
Shahin, Victor
author_facet Liashkovich, Ivan
Stefanello, Sílvio Terra
Vidyadharan, Reshma
Haufe, Günter
Erofeev, Alexander
Gorelkin, Peter V.
Kolmogorov, Vasilii
Mizdal, Caren Rigon
Dulebo, Alexander
Bulk, Etmar
Kouzel, Ian U.
Shahin, Victor
author_sort Liashkovich, Ivan
collection PubMed
description Clathrin‐mediated endocytosis (CME) is an essential cell physiological process of broad biomedical relevance. Since the recent introduction of Pitstop‐2 as a potent CME inhibitor, we and others have reported on substantial clathrin‐independent inhibitory effects. Herein, we developed and experimentally validated a novel fluorescent derivative of Pitstop‐2, termed RVD‐127, to clarify Pitstop‐2 diverse effects. Using RVD‐127, we were able to trace additional protein targets of Pitstop‐2. Besides inhibiting CME, Pitstop‐2 and RVD‐127 proved to directly and reversibly bind to at least two members of the small GTPase superfamily Ran and Rac1 with particularly high efficacy. Binding locks the GTPases in a guanosine diphosphate (GDP)‐like conformation disabling their interaction with their downstream effectors. Consequently, overall cell motility, mechanics and nucleocytoplasmic transport integrity are rapidly disrupted at inhibitor concentrations well below those required to significantly reduce CME. We conclude that Pitstop‐2 is a highly potent, reversible inhibitor of small GTPases. The inhibition of these molecular switches of diverse crucial signaling pathways, including nucleocytoplasmic transport and overall cell dynamics and motility, clarifies the diversity of Pitstop‐2 activities. Moreover, considering the fundamental importance and broad implications of small GTPases in physiology, pathophysiology and drug development, Pitstop‐2 and RVD‐127 open up novel avenues.
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spelling pubmed-103547672023-07-20 Pitstop‐2 and its novel derivative RVD‐127 disrupt global cell dynamics and nuclear pores integrity by direct interaction with small GTPases Liashkovich, Ivan Stefanello, Sílvio Terra Vidyadharan, Reshma Haufe, Günter Erofeev, Alexander Gorelkin, Peter V. Kolmogorov, Vasilii Mizdal, Caren Rigon Dulebo, Alexander Bulk, Etmar Kouzel, Ian U. Shahin, Victor Bioeng Transl Med Research Articles Clathrin‐mediated endocytosis (CME) is an essential cell physiological process of broad biomedical relevance. Since the recent introduction of Pitstop‐2 as a potent CME inhibitor, we and others have reported on substantial clathrin‐independent inhibitory effects. Herein, we developed and experimentally validated a novel fluorescent derivative of Pitstop‐2, termed RVD‐127, to clarify Pitstop‐2 diverse effects. Using RVD‐127, we were able to trace additional protein targets of Pitstop‐2. Besides inhibiting CME, Pitstop‐2 and RVD‐127 proved to directly and reversibly bind to at least two members of the small GTPase superfamily Ran and Rac1 with particularly high efficacy. Binding locks the GTPases in a guanosine diphosphate (GDP)‐like conformation disabling their interaction with their downstream effectors. Consequently, overall cell motility, mechanics and nucleocytoplasmic transport integrity are rapidly disrupted at inhibitor concentrations well below those required to significantly reduce CME. We conclude that Pitstop‐2 is a highly potent, reversible inhibitor of small GTPases. The inhibition of these molecular switches of diverse crucial signaling pathways, including nucleocytoplasmic transport and overall cell dynamics and motility, clarifies the diversity of Pitstop‐2 activities. Moreover, considering the fundamental importance and broad implications of small GTPases in physiology, pathophysiology and drug development, Pitstop‐2 and RVD‐127 open up novel avenues. John Wiley & Sons, Inc. 2022-10-19 /pmc/articles/PMC10354767/ /pubmed/37476059 http://dx.doi.org/10.1002/btm2.10425 Text en © 2022 The Authors. Bioengineering & Translational Medicine published by Wiley Periodicals LLC on behalf of American Institute of Chemical Engineers. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Liashkovich, Ivan
Stefanello, Sílvio Terra
Vidyadharan, Reshma
Haufe, Günter
Erofeev, Alexander
Gorelkin, Peter V.
Kolmogorov, Vasilii
Mizdal, Caren Rigon
Dulebo, Alexander
Bulk, Etmar
Kouzel, Ian U.
Shahin, Victor
Pitstop‐2 and its novel derivative RVD‐127 disrupt global cell dynamics and nuclear pores integrity by direct interaction with small GTPases
title Pitstop‐2 and its novel derivative RVD‐127 disrupt global cell dynamics and nuclear pores integrity by direct interaction with small GTPases
title_full Pitstop‐2 and its novel derivative RVD‐127 disrupt global cell dynamics and nuclear pores integrity by direct interaction with small GTPases
title_fullStr Pitstop‐2 and its novel derivative RVD‐127 disrupt global cell dynamics and nuclear pores integrity by direct interaction with small GTPases
title_full_unstemmed Pitstop‐2 and its novel derivative RVD‐127 disrupt global cell dynamics and nuclear pores integrity by direct interaction with small GTPases
title_short Pitstop‐2 and its novel derivative RVD‐127 disrupt global cell dynamics and nuclear pores integrity by direct interaction with small GTPases
title_sort pitstop‐2 and its novel derivative rvd‐127 disrupt global cell dynamics and nuclear pores integrity by direct interaction with small gtpases
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10354767/
https://www.ncbi.nlm.nih.gov/pubmed/37476059
http://dx.doi.org/10.1002/btm2.10425
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