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

• Published in: Acta crystallographica. Section D, Biological crystallography. Vol. 60; no. 11; pp. 2063 - 2066
• Main Authors: Newstead, Simon, Chien, Chin-Hsiang, Taylor, Margaret, Taylor, Garry
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England Munksgaard International Publishers 01.11.2004
• Subjects: Catalytic Domain; Clostridium perfringens - enzymology; Clostridium perfringens - genetics; Crystallization; Crystallography, X-Ray; nanI; Neuraminidase - chemistry; Neuraminidase - genetics; sialidases
• ISSN: 1399-0047; 0907-4449; 1399-0047
• DOI: 10.1107/S090744490402181X

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 P212121, with unit‐cell parameters a = 96.98, b = 69.41, c = 72.69 Å and one monomer per asymmetric unit. The crystals diffract to at least 0.92 Å. A molecular‐replacement solution was obtained using the catalytic domain of the sialidase from the leech Macrobdella decora.