Functional analysis of a group A streptococcal glycoside hydrolase Spy1600 from family 84 reveals it is a β-N-acetylglucosaminidase and not a hyaluronidase

Sheldon, William, Macauley, Matthew, Taylor, Edward, Robinson, Charlotte, Charnock, Simon, Davies, Gideon, Vocadlo, David and Black, Gary (2006) Functional analysis of a group A streptococcal glycoside hydrolase Spy1600 from family 84 reveals it is a β-N-acetylglucosaminidase and not a hyaluronidase. Biochemical Journal, 399 (2). pp. 241-247. ISSN 0264 6021

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Official URL: http://dx.doi.org/10.1042/BJ20060307

Abstract

Group A streptococcus (Streptococcus pyogenes) is the causative agent of severe invasive infections such as necrotizing fasciitis (the so-called 'flesh eating disease') and toxic-shock syndrome. Spy 1600, a glycoside hydrolase from family 84 of the large superfamily of glycoside hydrolases, has been proposed to be a virulence factor. In the present study we show that Spy1600 has no activity toward galactosaminides or hyaluronan, but does remove β-O-linked N-acetylglucosamine from mammalian glycoproteins - an observation consistent with the inclusion of eukaryotic O-glycoprotein 2-acetamido-2-deoxy-β-D- glucopyranosidases within glycoside hydrolase family 84. Proton NMR studies, structure-reactivity studies for a series of fluorinated analogues and analysis of 1,2-dideoxy-2′-methyl-α-D-glucopyranoso-[2,1-d]- Δ2′-thiazoline as a competitive inhibitor reveals that Spy1600 uses a double-displacement mechanism involving substrate-assisted catalysis. Family 84 glycoside hydrolases are therefore comprised of both prokaryotic and eukaryotic β-N-acetylglucosaminidases using a conserved catalytic mechanism involving substrate-assisted catalysis. Since these enzymes do not have detectable hyaluronidase activity, many family 84 glycoside hydrolases are most likely incorrectly annotated as hyaluronidases.

Item Type: Article
Uncontrolled Keywords: β-N-acetylglucosaminidase (GlcNAcase), 1,2-dideoxy-2′-methyl-α-D-glucopyranoso-[2,1-d]- Δ2′-thiazoline (NAG-thiazoline), mammalian glycoproteins
Subjects: F100 Chemistry
Department: Faculties > Health and Life Sciences > Applied Sciences
Depositing User: Becky Skoyles
Date Deposited: 30 Jan 2015 09:26
Last Modified: 24 Oct 2017 11:27
URI: http://nrl.northumbria.ac.uk/id/eprint/20046

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