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Structural and Kinetic Analysis of Bacillus subtilis N-Acetylglucosaminidase Reveals a Unique Asp-His Dyad Mechanism

Structural and Kinetic Analysis of Bacillus subtilis N-Acetylglucosaminidase Reveals a Unique Asp-His Dyad Mechanism

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LITZINGER, Silke, Stefanie FISCHER, Patrick POLZER, Kay DIEDERICHS, Wolfram WELTE, Christoph MAYER, 2010. Structural and Kinetic Analysis of Bacillus subtilis N-Acetylglucosaminidase Reveals a Unique Asp-His Dyad Mechanism. In: The Journal of Biological Chemistry. 285(46), pp. 35675-35684. ISSN 0021-9258. eISSN 1083-351X. Available under: doi: 10.1074/jbc.M110.131037

@article{Litzinger2010Struc-13554, title={Structural and Kinetic Analysis of Bacillus subtilis N-Acetylglucosaminidase Reveals a Unique Asp-His Dyad Mechanism}, year={2010}, doi={10.1074/jbc.M110.131037}, number={46}, volume={285}, issn={0021-9258}, journal={The Journal of Biological Chemistry}, pages={35675--35684}, author={Litzinger, Silke and Fischer, Stefanie and Polzer, Patrick and Diederichs, Kay and Welte, Wolfram and Mayer, Christoph} }

<rdf:RDF xmlns:dcterms="http://purl.org/dc/terms/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#" xmlns:bibo="http://purl.org/ontology/bibo/" xmlns:dspace="http://digital-repositories.org/ontologies/dspace/0.1.0#" xmlns:foaf="http://xmlns.com/foaf/0.1/" xmlns:void="http://rdfs.org/ns/void#" xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > <rdf:Description rdf:about="https://kops.uni-konstanz.de/rdf/resource/123456789/13554"> <dc:contributor>Welte, Wolfram</dc:contributor> <dc:contributor>Mayer, Christoph</dc:contributor> <foaf:homepage rdf:resource="http://localhost:8080/jspui"/> <dc:creator>Mayer, Christoph</dc:creator> <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-05-31T09:33:05Z</dc:date> <dc:language>eng</dc:language> <dc:creator>Polzer, Patrick</dc:creator> <dcterms:rights rdf:resource="http://nbn-resolving.org/urn:nbn:de:bsz:352-20140905103605204-4002607-1"/> <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-10-30T23:25:04Z</dcterms:available> <dc:creator>Fischer, Stefanie</dc:creator> <dcterms:abstract xml:lang="eng">Three-dimensional structures of NagZ of Bacillus subtilis, the first structures of a two-domain β-N-acetylglucosaminidase of family 3 of glycosidases, were determined with and without the transition state mimicking inhibitor PUGNAc bound to the active site, at 1.84- and 1.40-Å resolution, respectively. The structures together with kinetic analyses of mutants revealed an Asp-His dyad involved in catalysis: His234 of BsNagZ acts as general acid/base catalyst and is hydrogen bonded by Asp232 for proper function. Replacement of both His234 and Asp232 with glycine reduced the rate of hydrolysis of the fluorogenic substrate 4′-methylumbelliferyl N-acetyl-β-d-glucosaminide 1900- and 4500-fold, respectively, and rendered activity pH-independent in the alkaline range consistent with a role of these residues in acid/base catalysis. N-Acetylglucosaminyl enzyme intermediate accumulated in the H234G mutant and β-azide product was formed in the presence of sodium azide in both mutants. The Asp-His dyad is conserved within β-N-acetylglucosaminidases but otherwise absent in β-glycosidases of family 3, which instead carry a “classical” glutamate acid/base catalyst. The acid/base glutamate of Hordeum vulgare exoglucanase (Exo1) superimposes with His234 of the dyad of BsNagZ and, in contrast to the latter, protrudes from a second domain of the enzyme into the active site. This is the first report of an Asp-His catalytic dyad involved in hydrolysis of glycosides resembling in function the Asp-His-Ser triad of serine proteases. Our findings will facilitate the development of mechanism-based inhibitors that selectively target family 3 β-N-acetylglucosaminidases, which are involved in bacterial cell wall turnover, spore germination, and induction of β-lactamase.</dcterms:abstract> <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/rdf/resource/123456789/28"/> <dcterms:bibliographicCitation>First publ. in: The Journal of Biological Chemistry 285 (2010), 46, pp. 35675-35684, doi: 10.1074/jbc.M110.131037</dcterms:bibliographicCitation> <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/> <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/rdf/resource/123456789/28"/> <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/13554/2/J.%20Biol.%20Chem.-2010-Litzinger_Structural%20analysis.pdf"/> <dc:contributor>Fischer, Stefanie</dc:contributor> <dc:contributor>Diederichs, Kay</dc:contributor> <dc:creator>Diederichs, Kay</dc:creator> <dc:creator>Litzinger, Silke</dc:creator> <dc:creator>Welte, Wolfram</dc:creator> <dc:rights>deposit-license</dc:rights> <dcterms:title>Structural and Kinetic Analysis of Bacillus subtilis N-Acetylglucosaminidase Reveals a Unique Asp-His Dyad Mechanism</dcterms:title> <dc:contributor>Polzer, Patrick</dc:contributor> <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/13554/2/J.%20Biol.%20Chem.-2010-Litzinger_Structural%20analysis.pdf"/> <dcterms:issued>2010</dcterms:issued> <dc:contributor>Litzinger, Silke</dc:contributor> <bibo:uri rdf:resource="http://kops.uni-konstanz.de/handle/123456789/13554"/> </rdf:Description> </rdf:RDF>

Dateiabrufe seit 01.10.2014 (Informationen über die Zugriffsstatistik)

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