Folding energetics of a multidomain protein, flagellin

Shinya Honda, Hatsuho Uedaira, F. Vonderviszt, Shun Ichi Kidokoro, Keiichi Namba

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Thermodynamic investigations of flagellin from Salmonella typhimurium and its proteolytic fragments were conducted by differential scanning calorimetry (DSC) and circular dichroism (CD) melting measurements. A new method of analysis for a multi-state transition based on our original theoretical treatment of thermodynamic equations has been developed to analyze those data. The analysis of DSC curves confirmed the three thermodynamic domains of flagellin. The thermodynamic parameters of each domain were revised from those previously reported and the new values of the parameters have a good correlation to the apparent molecular masses of the morphological domains. CD melting measurements at far and near-UV wavelengths showed sequential unfolding of the domains. Therefore, we could reasonably assign the thermodynamically identified domains to the morphological domains. Further analysis of both DSC and CD data provided insights into the folding energetics of the multidomain structure of flagellin. An inner domain (D(f)1) of flagellin in the filament unfolds through a relatively broad transition, while the two outer domains unfold cooperatively and show sharp transitions. This indicates that the interdomain interactions between D(f)1 and D2 has different characteristics from the apparently more intimate interactions between D2 and D3. These characteristics suggest that flagellin is organized with relatively flexible domains and rigid domains, which appears to be responsible for the well-regulated assembly mechanism of the bacterial flagellar filament.

Original languageEnglish
Pages (from-to)719-732
Number of pages14
JournalJournal of Molecular Biology
Volume293
Issue number3
DOIs
Publication statusPublished - Oct 29 1999

Fingerprint

Flagellin
Thermodynamics
Differential Scanning Calorimetry
Circular Dichroism
Proteins
Freezing
Salmonella typhimurium

Keywords

  • Circular dichroism
  • Differential scanning calorimetry
  • Flagellin
  • Folding energetics
  • Multidomain protein

ASJC Scopus subject areas

  • Virology

Cite this

Folding energetics of a multidomain protein, flagellin. / Honda, Shinya; Uedaira, Hatsuho; Vonderviszt, F.; Kidokoro, Shun Ichi; Namba, Keiichi.

In: Journal of Molecular Biology, Vol. 293, No. 3, 29.10.1999, p. 719-732.

Research output: Contribution to journalArticle

Honda, Shinya ; Uedaira, Hatsuho ; Vonderviszt, F. ; Kidokoro, Shun Ichi ; Namba, Keiichi. / Folding energetics of a multidomain protein, flagellin. In: Journal of Molecular Biology. 1999 ; Vol. 293, No. 3. pp. 719-732.
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