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Tytuł:
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Delivery of antibody mimics into mammalian cells via anthrax toxin protective antigen.
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Autorzy:
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Liao X; Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue 18-596, Cambridge, MA 02193 (USA).
Rabideau AE
Pentelute BL
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Źródło:
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Chembiochem : a European journal of chemical biology [Chembiochem] 2014 Nov 03; Vol. 15 (16), pp. 2458-66. Date of Electronic Publication: 2014 Sep 22.
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Typ publikacji:
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Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.
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Język:
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English
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Imprint Name(s):
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Original Publication: Weinheim, Germany : Wiley-VCH Verlag, c2000-
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MeSH Terms:
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Antibodies/*metabolism
Antigens/*metabolism
Antigens, Bacterial/*chemistry
Bacterial Toxins/*chemistry
Biocompatible Materials/*chemistry
Animals ; Antibodies/chemistry ; Antigens/chemistry ; Biocompatible Materials/metabolism ; CHO Cells ; Cell-Penetrating Peptides/chemistry ; Cell-Penetrating Peptides/metabolism ; Cricetinae ; Cricetulus ; Fusion Proteins, bcr-abl/antagonists & inhibitors ; Fusion Proteins, bcr-abl/metabolism ; HEK293 Cells ; Humans ; Immunoprecipitation ; K562 Cells ; MAP Kinase Signaling System ; Protein Interaction Domains and Motifs ; Protein Structure, Tertiary ; Recombinant Fusion Proteins/genetics ; Recombinant Fusion Proteins/metabolism ; raf Kinases/antagonists & inhibitors ; raf Kinases/metabolism ; ras Proteins/antagonists & inhibitors ; ras Proteins/metabolism
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References:
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Grant Information:
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U54 AI057159 United States AI NIAID NIH HHS
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Contributed Indexing:
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Keywords: anthrax toxin; antibodies; intracellular delivery; protein-protein interactions; sortases
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Substance Nomenclature:
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0 (Antibodies)
0 (Antigens)
0 (Antigens, Bacterial)
0 (Bacterial Toxins)
0 (Biocompatible Materials)
0 (Cell-Penetrating Peptides)
0 (Recombinant Fusion Proteins)
0 (anthrax toxin)
EC 2.7.10.2 (Fusion Proteins, bcr-abl)
EC 2.7.11.1 (raf Kinases)
EC 3.6.5.2 (ras Proteins)
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Entry Date(s):
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Date Created: 20140925 Date Completed: 20150630 Latest Revision: 20231111
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Update Code:
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20240104
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PubMed Central ID:
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PMC4498471
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DOI:
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10.1002/cbic.201402290
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PMID:
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25250705
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Antibody mimics have significant scientific and therapeutic utility for the disruption of protein-protein interactions inside cells; however, their delivery to the cell cytosol remains a major challenge. Here we show that protective antigen (PA), a component of anthrax toxin, efficiently transports commonly used antibody mimics to the cytosol of mammalian cells when conjugated to the N-terminal domain of LF (LFN). In contrast, a cell-penetrating peptide (CPP) was not able to deliver any of these antibody mimics into the cell cytosol. The refolding and binding of a transported tandem monobody to Bcr-Abl (its protein target) in chronic myeloid leukemia cells were confirmed by co-immunoprecipitation. We also observed inhibition of Bcr-Abl kinase activity and induction of apoptosis caused by the monobody. In a separate case, we show disruption of key interactions in the MAPK signaling pathway after PA-mediated delivery of an affibody binder that targets hRaf-1. We show for the first time that PA can deliver bioactive antibody mimics to disrupt intracellular protein-protein interactions. This technology adds a useful tool to expand the applications of these modern agents to the intracellular milieu.
(© 2014 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the Creative Commons Attribution Non-Commercial NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.)