Structural and folding dynamic properties of the T70N variant of human lysozyme

Gennaro Esposito, Julian Garcia, Palma Mangione, Sofia Giorgetti, Alessandra Corazza, Paolo Viglino, Fabrizio Chiti, Alessia Andreola, Pascal Dumy, David Booth, Philip N. Hawkins, Vittorio Bellotti

    Research output: Contribution to journalArticle

    Abstract

    Definition of the transition mechanism from the native globular protein into fibrillar polymer was greatly improved by the biochemical and biophysical studies carried out on the two amyloidogenic variants of human lysozyme, I56T and D67H. Here we report thermodynamic and kinetic data on folding as well as structural features of a naturally occurring variant of human lysozyme, T70N, which is present in the British population at an allele frequency of 5% and, according to clinical and histopathological data, is not amyloidogenic. This variant is less stable than the wild-type protein by 3.7 kcal/mol, but more stable than the pathological, amyloidogenic variants. Unfolding kinetics in guanidine are six times faster than in the wild-type, but three and twenty times slower than in the amyloidogenic variants. Enzyme catalytic parameters, such as maximal velocity and affinity, are reduced in comparison to the wild-type. The solution structure, determined by 1H NMR and modeling calculations, exhibits a more compact arrangement at the interface between the β-sheet domain and the subsequent loop on one side and part of the α domain on the other side, compared with the wild-type protein. This is the opposite of the conformational variation shown by the amyloidogenic variant D67H, but it accounts for the reduced stability and catalytic performance of T70N.

    Original languageEnglish (US)
    Pages (from-to)25910-25918
    Number of pages9
    JournalJournal of Biological Chemistry
    Volume278
    Issue number28
    DOIs
    StatePublished - Jul 11 2003

    Fingerprint

    Muramidase
    Proteins
    Kinetics
    Guanidine
    Thermodynamics
    Gene Frequency
    Polymers
    Nuclear magnetic resonance
    Enzymes
    Population

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology
    • Cell Biology

    Cite this

    Esposito, G., Garcia, J., Mangione, P., Giorgetti, S., Corazza, A., Viglino, P., ... Bellotti, V. (2003). Structural and folding dynamic properties of the T70N variant of human lysozyme. Journal of Biological Chemistry, 278(28), 25910-25918. https://doi.org/10.1074/jbc.M211000200

    Structural and folding dynamic properties of the T70N variant of human lysozyme. / Esposito, Gennaro; Garcia, Julian; Mangione, Palma; Giorgetti, Sofia; Corazza, Alessandra; Viglino, Paolo; Chiti, Fabrizio; Andreola, Alessia; Dumy, Pascal; Booth, David; Hawkins, Philip N.; Bellotti, Vittorio.

    In: Journal of Biological Chemistry, Vol. 278, No. 28, 11.07.2003, p. 25910-25918.

    Research output: Contribution to journalArticle

    Esposito, G, Garcia, J, Mangione, P, Giorgetti, S, Corazza, A, Viglino, P, Chiti, F, Andreola, A, Dumy, P, Booth, D, Hawkins, PN & Bellotti, V 2003, 'Structural and folding dynamic properties of the T70N variant of human lysozyme', Journal of Biological Chemistry, vol. 278, no. 28, pp. 25910-25918. https://doi.org/10.1074/jbc.M211000200
    Esposito G, Garcia J, Mangione P, Giorgetti S, Corazza A, Viglino P et al. Structural and folding dynamic properties of the T70N variant of human lysozyme. Journal of Biological Chemistry. 2003 Jul 11;278(28):25910-25918. https://doi.org/10.1074/jbc.M211000200
    Esposito, Gennaro ; Garcia, Julian ; Mangione, Palma ; Giorgetti, Sofia ; Corazza, Alessandra ; Viglino, Paolo ; Chiti, Fabrizio ; Andreola, Alessia ; Dumy, Pascal ; Booth, David ; Hawkins, Philip N. ; Bellotti, Vittorio. / Structural and folding dynamic properties of the T70N variant of human lysozyme. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 28. pp. 25910-25918.
    @article{9c03f68464744c348fb96f82455b48cb,
    title = "Structural and folding dynamic properties of the T70N variant of human lysozyme",
    abstract = "Definition of the transition mechanism from the native globular protein into fibrillar polymer was greatly improved by the biochemical and biophysical studies carried out on the two amyloidogenic variants of human lysozyme, I56T and D67H. Here we report thermodynamic and kinetic data on folding as well as structural features of a naturally occurring variant of human lysozyme, T70N, which is present in the British population at an allele frequency of 5{\%} and, according to clinical and histopathological data, is not amyloidogenic. This variant is less stable than the wild-type protein by 3.7 kcal/mol, but more stable than the pathological, amyloidogenic variants. Unfolding kinetics in guanidine are six times faster than in the wild-type, but three and twenty times slower than in the amyloidogenic variants. Enzyme catalytic parameters, such as maximal velocity and affinity, are reduced in comparison to the wild-type. The solution structure, determined by 1H NMR and modeling calculations, exhibits a more compact arrangement at the interface between the β-sheet domain and the subsequent loop on one side and part of the α domain on the other side, compared with the wild-type protein. This is the opposite of the conformational variation shown by the amyloidogenic variant D67H, but it accounts for the reduced stability and catalytic performance of T70N.",
    author = "Gennaro Esposito and Julian Garcia and Palma Mangione and Sofia Giorgetti and Alessandra Corazza and Paolo Viglino and Fabrizio Chiti and Alessia Andreola and Pascal Dumy and David Booth and Hawkins, {Philip N.} and Vittorio Bellotti",
    year = "2003",
    month = "7",
    day = "11",
    doi = "10.1074/jbc.M211000200",
    language = "English (US)",
    volume = "278",
    pages = "25910--25918",
    journal = "Journal of Biological Chemistry",
    issn = "0021-9258",
    publisher = "American Society for Biochemistry and Molecular Biology Inc.",
    number = "28",

    }

    TY - JOUR

    T1 - Structural and folding dynamic properties of the T70N variant of human lysozyme

    AU - Esposito, Gennaro

    AU - Garcia, Julian

    AU - Mangione, Palma

    AU - Giorgetti, Sofia

    AU - Corazza, Alessandra

    AU - Viglino, Paolo

    AU - Chiti, Fabrizio

    AU - Andreola, Alessia

    AU - Dumy, Pascal

    AU - Booth, David

    AU - Hawkins, Philip N.

    AU - Bellotti, Vittorio

    PY - 2003/7/11

    Y1 - 2003/7/11

    N2 - Definition of the transition mechanism from the native globular protein into fibrillar polymer was greatly improved by the biochemical and biophysical studies carried out on the two amyloidogenic variants of human lysozyme, I56T and D67H. Here we report thermodynamic and kinetic data on folding as well as structural features of a naturally occurring variant of human lysozyme, T70N, which is present in the British population at an allele frequency of 5% and, according to clinical and histopathological data, is not amyloidogenic. This variant is less stable than the wild-type protein by 3.7 kcal/mol, but more stable than the pathological, amyloidogenic variants. Unfolding kinetics in guanidine are six times faster than in the wild-type, but three and twenty times slower than in the amyloidogenic variants. Enzyme catalytic parameters, such as maximal velocity and affinity, are reduced in comparison to the wild-type. The solution structure, determined by 1H NMR and modeling calculations, exhibits a more compact arrangement at the interface between the β-sheet domain and the subsequent loop on one side and part of the α domain on the other side, compared with the wild-type protein. This is the opposite of the conformational variation shown by the amyloidogenic variant D67H, but it accounts for the reduced stability and catalytic performance of T70N.

    AB - Definition of the transition mechanism from the native globular protein into fibrillar polymer was greatly improved by the biochemical and biophysical studies carried out on the two amyloidogenic variants of human lysozyme, I56T and D67H. Here we report thermodynamic and kinetic data on folding as well as structural features of a naturally occurring variant of human lysozyme, T70N, which is present in the British population at an allele frequency of 5% and, according to clinical and histopathological data, is not amyloidogenic. This variant is less stable than the wild-type protein by 3.7 kcal/mol, but more stable than the pathological, amyloidogenic variants. Unfolding kinetics in guanidine are six times faster than in the wild-type, but three and twenty times slower than in the amyloidogenic variants. Enzyme catalytic parameters, such as maximal velocity and affinity, are reduced in comparison to the wild-type. The solution structure, determined by 1H NMR and modeling calculations, exhibits a more compact arrangement at the interface between the β-sheet domain and the subsequent loop on one side and part of the α domain on the other side, compared with the wild-type protein. This is the opposite of the conformational variation shown by the amyloidogenic variant D67H, but it accounts for the reduced stability and catalytic performance of T70N.

    UR - http://www.scopus.com/inward/record.url?scp=0037830693&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=0037830693&partnerID=8YFLogxK

    U2 - 10.1074/jbc.M211000200

    DO - 10.1074/jbc.M211000200

    M3 - Article

    VL - 278

    SP - 25910

    EP - 25918

    JO - Journal of Biological Chemistry

    JF - Journal of Biological Chemistry

    SN - 0021-9258

    IS - 28

    ER -