Nucleophosmin-1 regions associated with acute myeloid leukemia interact differently with lipid membranes |
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Authors: | Augusta De Santis Sara La Manna Irene Russo Krauss Anna Maria Malfitano Ettore Novellino Luca Federici Antonella De Cola Adele Di Matteo Gerardino DErrico Daniela Marasco |
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Institution: | 1. Department of Chemical Sciences, University of Naples “Federico II”, Naples, Italy;2. CSGI – Consorzio Interuniversitario per lo Sviluppo dei Sistemi a Grande Interfase, Florence, Italy;3. Department of Pharmacy, CIRPEB: Centro Interuniversitario di Ricerca sui Peptidi Bioattivi, University of Naples “Federico II”, 80134, Naples, Italy;4. Department of Medical, Oral and Biotechnological Sciences and CeSI-MeT, University of Chieti “G. d''Annunzio”, Via dei Vestini 31, 66100 Chieti, Italy;5. Institute of Molecular Biology and Pathology, CNR, Piazzale Aldo Moro 5, 00185 Rome, Italy |
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Abstract: | BackgroundNucleophosmin-1 (NPM1) is an abundant multifunctional protein, implicated in a variety of biological processes and in the pathogenesis of several human malignancies. Its C-terminal domain (CTD) is endowed with a three helix bundle and we demonstrated that several regions within it, associated with acute myeloid leukemia (AML), have a strong tendency to form beta amyloid-like assemblies toxic for cells. The central helix of the bundle (H2) resulted the most amyloidgenic region; here we aim to model the cytoxicity processes of the H2 sequence and getting clues of a potential involvement in toxicity of the interaction between CTDs and cellular membranes.MethodsWe investigated the interaction of CTD-NPM1 regions with model membranes through fluorescence, SPR, CD and ESR spectroscopies and the localization of NPM1 by immune-fluorescence in leukemic cells.ResultsOur findings indicate that investigated regions are able to interact with membranes with different mechanisms and outlined the importance of the presence of cholesterol.ConclusionsH2 showed a preference of interaction with membrane containing cholesterol determining a sensitive fluidification of the bilayer, while N-term H2 causes a stiffening of central and outer regions of the lipid system. Noticeably, NPM1 mut A demonstrated to thicken at the plasma membrane, differently from wt. These findings were corroborated by diverse mechanisms of interaction of CTDs toward membrane models in vitro.General significanceThis study suggests that the direct interaction of several regions of NPM1CTD with cellular membranes could be implicated in diseases where NPM1 is mutated and/or where its overexpression is cytoxic. |
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Keywords: | LC–MS Liquid Chromatography Mass Spectrometry CD circular dichroism ESR Electron Spin Resonance SPR Surface Plasmon Resonance CTD C-terminal domain NPM1 Nucleophosmin LUV Large Unilamellar Vesicle AML acute myeloid leukemia HFIP (1 1 1 3 3 3-Hexafluoro-2-propanol) ANS (8-anilinonaphthalene-1-sulfonate ammonium salt) PCSL Phosphocholine spin-label CNO 25-Doxyl-cholesterol CD spectroscopy ESR spectroscopy Protein-membrane interaction Nucleophosmin C-terminal domain |
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