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

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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, 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.
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