Comparative structural analysis of low-molecular mass fragments of Rapana venosa hemocyanin obtained using two different procedures

Annalaura Sabatucci, Patrice Vachette, Mariano Beltramini, Benedetto Salvato, Enrico Dainese

Research output: Contribution to journalArticlepeer-review

Abstract

Different fragments of the hemocyanin (Hc) isolated from the gastropod Rapana venosa containing a single functional unit (50 kDa), two functional units (100 kDa) and three functional units (150 kDa) were obtained in a dissociating buffer in the presence of Zn2+ and purified to homogeneity. Their conformations in solution were studied by means of small angle X-ray scattering (SAXS) and compared with those of the corresponding fragments previously obtained by limited proteolysis [Arch. Biochem. Biophys., 2000, 373, 154]. The overall shape of each fragment was determined using an ab initio approach. The crystal structures of the functional unit e from the same Hc and from another molluscan Hc (Octopus dofleini) were used to model 100 and 150 kDa fragments using rigid body movements to fit the corresponding SAXS patterns. Interesting differences were observed between the functional unit organization in the low-molecular mass fragments according to the two preparation methods, suggesting different localizations within the 11S functional subunit.

Original languageEnglish
Pages (from-to)127-137
Number of pages11
JournalJournal of Structural Biology
Volume149
Issue number2
DOIs
Publication statusPublished - Feb 2005

Keywords

  • Conformation
  • Functional unit
  • Hc
  • Low-resolution models
  • Proteolysis
  • SAXS
  • Size exclusion chromatography

ASJC Scopus subject areas

  • Structural Biology

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