Please use this identifier to cite or link to this item: http://hdl.handle.net/20.500.12779/5616
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dc.contributor.authorM., DEL VECCHIOen_us
dc.contributor.authorPogni, Rebeccaen_us
dc.contributor.authorBaratto, Maria Camillaen_us
dc.contributor.authorA., Nobbsen_us
dc.contributor.authorR., Rappuolien_us
dc.contributor.authorM., Pizzaen_us
dc.contributor.authorE., Balduccien_us
dc.date.accessioned2021-03-30T15:53:53Z-
dc.date.available2021-03-30T15:53:53Z-
dc.date.issued2009-
dc.identifier.issn0021-9258en_US
dc.identifier.urihttp://hdl.handle.net/20.500.12779/5616-
dc.description35794en_US
dc.description.abstractIn prokaryotes, mono ADP-ribose transfer enzymes represent a family of exotoxins that display activity in a variety of bacterial pathogens responsible for causing disease in plants and animals, including those affecting mankind such as diphtheria, cholera and whooping cough. We report here that NarE, a putative ADP-ribosylating toxin previously identified from Neisseria meningitidis, which shares structural homologies with Escherichia coli heat labile enterotoxin and toxin from Vibrio cholerae, possesses an iron-sulfur centre. The recombinant protein was expressed in E. coli and when purified at high concentration NarE has a distinctive golden brown in color. Evidence from UV-visible spectrophotometry and EPR spectroscopy revealed characteristics consistent of an iron-binding protein. The presence of iron was determined by colorimetric method and by Atomic Absorption Spectrophotometer. To obtain information about the identity of the amino-acids involved in binding iron, a combination of site-directed mutagenesis, UV-vis and enzymatic assays were performed. All four cysteine residues were individually replaced by serine. Substitution of C67 and C128 into serine caused a drastic reduction in the UV-vis signal intensity and in the A420/A280 ratio, suggesting that these two residues interact directly with the cluster and are essential for the formation of a stable coordination. This modification led to a consistent loss in ADP-ribosyltransferase activity while decrease in NAD-glycohydrolase was less dramatic in the C67S and C128S mutants indicating that the correct assembly of the iron-binding site is essential for exploiting transferase but not hydrolase activity. This is the first observation that a member of the ADP-ribosyltransferase family is an iron dependent enzyme and that contains a Fe-S cluster implicated in catalysis. This novel discovery may lead to the discovery of novel functions exerted by this class of enzymes.en_US
dc.language.isoenen_US
dc.relationNoneen_US
dc.relation.ispartofTHE JOURNAL OF BIOLOGICAL CHEMISTRYen_US
dc.titleIdentification of an Iron-sulfur Cluster that modulates the Enzymatic Activity in NarE, a Neisseria Meningitidis ADP-ribosyltransferaseen_US
dc.typeArticleen_US
dc.identifier.doi10.1074/jbc.M109.057547en_US
dc.identifier.pmid19744927en_US
dc.identifier.scopus2-s2.0-70450270635en_US
dc.identifier.isiWOS:000272028500006en_US
dc.relation.volume284en_US
dc.relation.issue48en_US
dc.description.firstpage33040en_US
dc.description.lastpage33047en_US
dc.description.thirdmissionNot applicableen_US
item.cerifentitytypePublications-
item.grantfulltextnone-
item.languageiso639-1en-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.openairetypeArticle-
item.fulltextNo Fulltext-
crisitem.author.orcid0000-0001-6681-1592-
crisitem.author.orcid0000-0003-2258-779X-
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