Effect of tetracyclines on the dynamics of formation and destructuration of β2-microglobulin amyloid fibrils

Sofia Giorgetti, Sara Raimondi, Katiuscia Pagano, Annalisa Relini, Monica Bucciantini, Alessandra Corazza, Federico Fogolari, Luca Codutti, Mario Salmona, Palma Mangione, Lino Colombo, Ada De Luigi, Riccardo Porcari, Alessandra Gliozzi, Massimo Stefani, Gennaro Esposito, Vittorio Bellotti, Monica Stoppini

    Research output: Contribution to journalArticle

    Abstract

    The discovery of methods suitable for the conversion in vitro of native proteins into amyloid fibrils has shed light on the molecular basis of amyloidosis and has provided fundamental tools for drug discovery. We have studied the capacity of a small library of tetracycline analogues to modulate the formation or destructuration of β2-microglobulin fibrils. The inhibition of fibrillogenesis of the wild type protein was first established in the presence of 20% trifluoroethanol and confirmed under a more physiologic environment including heparin and collagen. The latter conditions were also used to study the highly amyloidogenic variant, P32G. The NMR analysis showed that doxycycline inhibits β2-microglobulin self-association and stabilizes the native-like species through fast exchange interactions involving specific regions of the protein. Cell viability assays demonstrated that the drug abolishes the natural cytotoxic activity of soluble β2-microglobulin, further strengthening a possible in vivo therapeutic exploitation of this drug. Doxycycline can disassemble preformed fibrils, but the IC50 is 5-fold higher than that necessary for the inhibition of fibrillogenesis. Fibril destructuration is a dynamic and timedependent process characterized by the early formation of cytotoxic protein aggregates that, in a few hours, convert into non-toxic insoluble material. The efficacy of doxycycline as a drug against dialysis-related amyloidosis would benefit from the ability of the drug to accumulate just in the skeletal system where amyloid is formed. In these tissues, the doxycycline concentration reaches values several folds higher than those resulting in inhibition of amyloidogenesis and amyloid destructuration in vitro.

    Original languageEnglish (US)
    Pages (from-to)2121-2131
    Number of pages11
    JournalJournal of Biological Chemistry
    Volume286
    Issue number3
    DOIs
    StatePublished - Jan 21 2011

    Fingerprint

    Tetracyclines
    Doxycycline
    Amyloid
    Amyloidosis
    Pharmaceutical Preparations
    Trifluoroethanol
    Proteins
    Dialysis
    Exchange interactions
    Drug Discovery
    Tetracycline
    Inhibitory Concentration 50
    Heparin
    Assays
    Cell Survival
    Collagen
    Cells
    Nuclear magnetic resonance
    Association reactions
    Tissue

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology
    • Cell Biology

    Cite this

    Giorgetti, S., Raimondi, S., Pagano, K., Relini, A., Bucciantini, M., Corazza, A., ... Stoppini, M. (2011). Effect of tetracyclines on the dynamics of formation and destructuration of β2-microglobulin amyloid fibrils. Journal of Biological Chemistry, 286(3), 2121-2131. https://doi.org/10.1074/jbc.M110.178376

    Effect of tetracyclines on the dynamics of formation and destructuration of β2-microglobulin amyloid fibrils. / Giorgetti, Sofia; Raimondi, Sara; Pagano, Katiuscia; Relini, Annalisa; Bucciantini, Monica; Corazza, Alessandra; Fogolari, Federico; Codutti, Luca; Salmona, Mario; Mangione, Palma; Colombo, Lino; De Luigi, Ada; Porcari, Riccardo; Gliozzi, Alessandra; Stefani, Massimo; Esposito, Gennaro; Bellotti, Vittorio; Stoppini, Monica.

    In: Journal of Biological Chemistry, Vol. 286, No. 3, 21.01.2011, p. 2121-2131.

    Research output: Contribution to journalArticle

    Giorgetti, S, Raimondi, S, Pagano, K, Relini, A, Bucciantini, M, Corazza, A, Fogolari, F, Codutti, L, Salmona, M, Mangione, P, Colombo, L, De Luigi, A, Porcari, R, Gliozzi, A, Stefani, M, Esposito, G, Bellotti, V & Stoppini, M 2011, 'Effect of tetracyclines on the dynamics of formation and destructuration of β2-microglobulin amyloid fibrils', Journal of Biological Chemistry, vol. 286, no. 3, pp. 2121-2131. https://doi.org/10.1074/jbc.M110.178376
    Giorgetti, Sofia ; Raimondi, Sara ; Pagano, Katiuscia ; Relini, Annalisa ; Bucciantini, Monica ; Corazza, Alessandra ; Fogolari, Federico ; Codutti, Luca ; Salmona, Mario ; Mangione, Palma ; Colombo, Lino ; De Luigi, Ada ; Porcari, Riccardo ; Gliozzi, Alessandra ; Stefani, Massimo ; Esposito, Gennaro ; Bellotti, Vittorio ; Stoppini, Monica. / Effect of tetracyclines on the dynamics of formation and destructuration of β2-microglobulin amyloid fibrils. In: Journal of Biological Chemistry. 2011 ; Vol. 286, No. 3. pp. 2121-2131.
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