Cloning, expression, purification, crystallization and preliminary X‐ray diffraction studies of N‐acetylneuraminate lyase from methicillin‐resistant Staphylococcus aureus. Issue 3 (24th March 2013)

• Record Type: Journal Article
• Authors: North, Rachel A.; Kessans, Sarah A.; Atkinson, Sarah C.; Suzuki, Hironori; Watson, Andrew J. A.; Burgess, Benjamin R.; Angley, Lauren M.; Hudson, André O.; Varsani, Arvind; Griffin, Michael D. W.; Fairbanks, Antony J.; Dobson, Renwick C. J.
• Is Part Of: Acta crystallographica. Volume 69:Issue 3(2013:Mar.)
• Page Start: 306
• Page End: 312
• Publication Date: 2013-03-24
• Subjects: Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S1744309113003060
• Languages: English
• ISSNs: 1744-3091

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>The enzyme <italic>N</italic>‐acetylneuraminate lyase (EC 4.1.3.3) is involved in the metabolism of sialic acids. Specifically, the enzyme catalyzes the retro‐aldol cleavage of <italic>N</italic>‐acetylneuraminic acid to form <italic>N</italic>‐acetyl‐D‐mannosamine and pyruvate. Sialic acids comprise a large family of nine‐carbon amino sugars, all of which are derived from the parent compound <italic>N</italic>‐acetylneuraminic acid. In recent years, <italic>N</italic>‐acetylneuraminate lyase has received considerable attention from both mechanistic and structural viewpoints and has been recognized as a potential antimicrobial drug target. The <italic>N</italic>‐acetylneuraminate lyase gene was cloned from methicillin‐resistant <italic>Staphylococcus aureus</italic> genomic DNA, and recombinant protein was expressed and purified from <italic>Escherichia coli</italic> BL21 (DE3). The enzyme crystallized in a number of crystal forms, predominantly from PEG precipitants, with the best crystal diffracting to beyond 1.70 Å resolution in space group <italic>P</italic>2<sub>1</sub>. Molecular replacement indicates the presence of eight monomers per asymmetric unit. Understanding the structural biology of <italic>N</italic>‐acetylneuraminate lyase in pathogenic bacteria, such as methicillin‐resistant <italic>S. aureus</italic>, will provide insights for the development of future antimicrobials.</p> </abstract>