Variants of β2-microglobulin cleaved at lysine-58 retain the main conformational features of the native protein but are more conformationally heterogeneous and unstable at physiological temperature

Maria C. Mimmi, Thomas J.D. Jørgensen, Fabio Pettirossi, Alessandra Corazza, Paolo Viglino, Gennaro Esposito, Ersilia De Lorenzi, Sofia Giorgetti, Mette Pries, Dorthe B. Corlin, Mogens H. Nissen, Niels H.H. Heegaard

    Research output: Contribution to journalArticle

    Abstract

    Cleavage of the small amyloidogenic protein β2- microglobulin after lysine-58 renders it more prone to unfolding and aggregation. This is important for dialysis-related β2- microglobulin amyloidosis, since elevated levels of cleaved β2- microglobulin may be found in the circulation of dialysis patients. However, the solution structures of these cleaved β2-microglobulin variants have not yet been assessed using single-residue techniques. We here use such methods to examine β2-microglobulin cleaved after lysine-58 and the further processed variant (found in vivo) from which lysine-58 is removed. We find that the solution stability of both variants, especially of β2-microglobulin from which lysine-58 is removed, is much reduced compared to wild-type β2-microglobulin and is strongly dependent on temperature and protein concentration. 1H-NMR spectroscopy and amide hydrogen (1H/2H) exchange monitored by MS show that the overall three-dimensional structure of the variants is similar to that of wild-type β2-microglobulin at subphysiological temperatures. However, deviations do occur, especially in the arrangement of the B, D and E β-strands close to the D-E loop cleavage site at lysine-58, and the experiments suggest conformational heterogeneity of the two variants. Two-dimensional NMR spectroscopy indicates that this heterogeneity involves an equilibrium between the native-like fold and at least one conformational intermediate resembling intermediates found in other structurally altered β2-microglobulin molecules. This is the first single-residue resolution study of a specific β2-microglobulin variant that has been found circulating in dialysis patients. The instability and conformational heterogeneity of this variant suggest its involvement in β2-microglobulin amyloidogenicity in vivo.

    Original languageEnglish (US)
    Pages (from-to)2461-2474
    Number of pages14
    JournalFEBS Journal
    Volume273
    Issue number11
    DOIs
    StatePublished - Jun 1 2006

    Fingerprint

    Lysine
    Dialysis
    Temperature
    Proteins
    Nuclear magnetic resonance spectroscopy
    Magnetic Resonance Spectroscopy
    Amyloidogenic Proteins
    Amyloidosis
    Amides
    Hydrogen
    Agglomeration
    Molecules
    Experiments

    Keywords

    • Amyloidosis
    • Cleaved β-microglobulin
    • Human β-microglobulin
    • NMR
    • Protein conformation

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology
    • Cell Biology

    Cite this

    Variants of β2-microglobulin cleaved at lysine-58 retain the main conformational features of the native protein but are more conformationally heterogeneous and unstable at physiological temperature. / Mimmi, Maria C.; Jørgensen, Thomas J.D.; Pettirossi, Fabio; Corazza, Alessandra; Viglino, Paolo; Esposito, Gennaro; De Lorenzi, Ersilia; Giorgetti, Sofia; Pries, Mette; Corlin, Dorthe B.; Nissen, Mogens H.; Heegaard, Niels H.H.

    In: FEBS Journal, Vol. 273, No. 11, 01.06.2006, p. 2461-2474.

    Research output: Contribution to journalArticle

    Mimmi, MC, Jørgensen, TJD, Pettirossi, F, Corazza, A, Viglino, P, Esposito, G, De Lorenzi, E, Giorgetti, S, Pries, M, Corlin, DB, Nissen, MH & Heegaard, NHH 2006, 'Variants of β2-microglobulin cleaved at lysine-58 retain the main conformational features of the native protein but are more conformationally heterogeneous and unstable at physiological temperature', FEBS Journal, vol. 273, no. 11, pp. 2461-2474. https://doi.org/10.1111/j.1742-4658.2006.05254.x
    Mimmi, Maria C. ; Jørgensen, Thomas J.D. ; Pettirossi, Fabio ; Corazza, Alessandra ; Viglino, Paolo ; Esposito, Gennaro ; De Lorenzi, Ersilia ; Giorgetti, Sofia ; Pries, Mette ; Corlin, Dorthe B. ; Nissen, Mogens H. ; Heegaard, Niels H.H. / Variants of β2-microglobulin cleaved at lysine-58 retain the main conformational features of the native protein but are more conformationally heterogeneous and unstable at physiological temperature. In: FEBS Journal. 2006 ; Vol. 273, No. 11. pp. 2461-2474.
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    abstract = "Cleavage of the small amyloidogenic protein β2- microglobulin after lysine-58 renders it more prone to unfolding and aggregation. This is important for dialysis-related β2- microglobulin amyloidosis, since elevated levels of cleaved β2- microglobulin may be found in the circulation of dialysis patients. However, the solution structures of these cleaved β2-microglobulin variants have not yet been assessed using single-residue techniques. We here use such methods to examine β2-microglobulin cleaved after lysine-58 and the further processed variant (found in vivo) from which lysine-58 is removed. We find that the solution stability of both variants, especially of β2-microglobulin from which lysine-58 is removed, is much reduced compared to wild-type β2-microglobulin and is strongly dependent on temperature and protein concentration. 1H-NMR spectroscopy and amide hydrogen (1H/2H) exchange monitored by MS show that the overall three-dimensional structure of the variants is similar to that of wild-type β2-microglobulin at subphysiological temperatures. However, deviations do occur, especially in the arrangement of the B, D and E β-strands close to the D-E loop cleavage site at lysine-58, and the experiments suggest conformational heterogeneity of the two variants. Two-dimensional NMR spectroscopy indicates that this heterogeneity involves an equilibrium between the native-like fold and at least one conformational intermediate resembling intermediates found in other structurally altered β2-microglobulin molecules. This is the first single-residue resolution study of a specific β2-microglobulin variant that has been found circulating in dialysis patients. The instability and conformational heterogeneity of this variant suggest its involvement in β2-microglobulin amyloidogenicity in vivo.",
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    AU - Mimmi, Maria C.

    AU - Jørgensen, Thomas J.D.

    AU - Pettirossi, Fabio

    AU - Corazza, Alessandra

    AU - Viglino, Paolo

    AU - Esposito, Gennaro

    AU - De Lorenzi, Ersilia

    AU - Giorgetti, Sofia

    AU - Pries, Mette

    AU - Corlin, Dorthe B.

    AU - Nissen, Mogens H.

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    KW - Cleaved β-microglobulin

    KW - Human β-microglobulin

    KW - NMR

    KW - Protein conformation

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