Crystallisation and atomic resolution X-ray diffraction of the catalytic domain of the large sialidase, nanI, from Clostridium perfringens.

S Newstead, C-H Chien, M Taylor, Garry Lindsay Taylor

Research output: Contribution to journalArticlepeer-review

12 Citations (Scopus)

Abstract

Sialidases catalyse the removal of terminal sialic acids from a range of glycoproteins, glycolipids and oligosaccharides. They have been found in bacteria, viruses and parasites, where they play important roles in pathogenesis and/or microbial nutrition, and in mammalian cells, where they modulate cell-surface glycosylation associated with a range of cellular activities. Clostridium perfringens, a causative agent of gas gangrene and peritonitis in humans, possesses three sialidases: nanH, nanI and nanJ, with molecular weights of 42, 77 and 129 kDa, respectively. The two larger enzymes are secreted by the bacterium and are involved in the pathogenesis and nutrition of Clostridium. As part of a study to examine the structures of all three enzymes, crystallization of the 77 kDa nanI isoenzyme was attempted. The expressed full-length protein was found to degrade easily; a stable 50 kDa catalytic domain was therefore subcloned. This domain was overexpressed in Escherichia coli and produced crystals belonging to space group P2(1)2(1)2(1), with unit-cell parameters a=96.98, b=69.41, c=72.69 Angstrom and one monomer per asymmetric unit. The crystals diffract to at least 0.92 Angstrom. A molecular-replacement solution was obtained using the catalytic domain of the sialidase from the leech Macrobdella decora.

Original languageEnglish
Pages (from-to)2063-2066
Number of pages4
JournalActa Crystallographica. Section D, Biological crystallography
VolumeD60
DOIs
Publication statusPublished - Nov 2004

Keywords

  • CRUZI TRANS-SIALIDASE
  • CRYSTAL-STRUCTURE
  • INFLUENZA-VIRUS
  • HEMAGGLUTININ-NEURAMINIDASE
  • BACTERIAL SIALIDASES
  • EXPRESSION
  • INHIBITORS
  • ENZYME
  • FOLD
  • MODE

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