Tetrameric Structure of Thermostable Direct Hemolysin from Vibrio parahaemolyticus Revealed by Ultracentrifugation, Small-angle X-ray Scattering and Electron Microscopy

Daizo Hamada, Takashi Higurashi, Kouta Mayanagi, Tomoko Miyata, Takashi Fukui, Tatsuya Iida, Takeshi Honda, Itaru Yanagihara

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

The thermostable direct hemolysin (TDH) is a major virulence factor of Vibrio parahaemolyticus. We have characterized the conformational properties of TDH by small-angle X-ray scattering (SAXS), ultracentrifugation and transmission electron microscopy. Sedimentation equilibrium and velocity studies revealed that the protein is tetrameric in aqueous solvents. The Guinier plot derived from SAXS data provided a radius of gyration of 29.0 Å. The elongated pattern with a shoulder of a pair distance distribution function derived from SAXS data suggested the presence of molecules with an anisotropic shape having a maximum diameter of 98 Å. Electron microscopic image analysis of the negatively stained TDH oligomer showed the presence of C4 symmetric particles with edge and diagonal lengths of 65 Å and 80 Å, respectively. Shape reconstruction was carried out by ab initio calculations using the SAXS data with a C4 symmetric approximation. These results suggested that the tetrameric TDH assumes an oblate structure. The hydrodynamic parameters predicted from the ab initio model differed slightly from the experimental values, suggesting the presence of flexible segments.

Original languageEnglish
Pages (from-to)187-195
Number of pages9
JournalJournal of Molecular Biology
Volume365
Issue number1
DOIs
Publication statusPublished - Jan 5 2007

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

Fingerprint Dive into the research topics of 'Tetrameric Structure of Thermostable Direct Hemolysin from Vibrio parahaemolyticus Revealed by Ultracentrifugation, Small-angle X-ray Scattering and Electron Microscopy'. Together they form a unique fingerprint.

  • Cite this