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dc.contributor.authorRaasakka, Arneen_US
dc.contributor.authorMyllykoski, Mattien_US
dc.contributor.authorLaulumaa, Saaraen_US
dc.contributor.authorLehtimäki, Marien_US
dc.contributor.authorHärtlein, Michaelen_US
dc.contributor.authorMoulin, Martineen_US
dc.contributor.authorKursula, Inarien_US
dc.contributor.authorKursula, Petrien_US
dc.date.accessioned2016-08-01T12:40:14Z
dc.date.available2016-08-01T12:40:14Z
dc.date.issued2015-11-13
dc.PublishedScientific Reports 2015, 5:16520eng
dc.identifier.issn2045-2322
dc.identifier.urihttps://hdl.handle.net/1956/12373
dc.description.abstract2′,3′-cyclic nucleotide 3′-phosphodiesterase (CNPase) is an enzyme highly abundant in the central nervous system myelin of terrestrial vertebrates. The catalytic domain of CNPase belongs to the 2H phosphoesterase superfamily and catalyzes the hydrolysis of nucleoside 2′,3′-cyclic monophosphates to nucleoside 2′-monophosphates. The detailed reaction mechanism and the essential catalytic amino acids involved have been described earlier, but the roles of many amino acids in the vicinity of the active site have remained unknown. Here, several CNPase catalytic domain mutants were studied using enzyme kinetics assays, thermal stability experiments, and X-ray crystallography. Additionally, the crystal structure of a perdeuterated CNPase catalytic domain was refined at atomic resolution to obtain a detailed view of the active site and the catalytic mechanism. The results specify determinants of ligand binding and novel essential residues required for CNPase catalysis. For example, the aromatic side chains of Phe235 and Tyr168 are crucial for substrate binding, and Arg307 may affect active site electrostatics and regulate loop dynamics. The β5-α7 loop, unique for CNPase in the 2H phosphoesterase family, appears to have various functions in the CNPase reaction mechanism, from coordinating the nucleophilic water molecule to providing a binding pocket for the product and being involved in product release.en_US
dc.language.isoengeng
dc.publisherNature Publishing Groupeng
dc.rightsAttribution CC BYeng
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/eng
dc.titleDeterminants of ligand binding and catalytic activity in the myelin enzyme 2′,3′-cyclic nucleotide 3′-phosphodiesteraseen_US
dc.typePeer reviewed
dc.typeJournal article
dc.date.updated2016-04-11T09:16:25Z
dc.description.versionpublishedVersionen_US
dc.identifier.doihttps://doi.org/10.1038/srep16520
dc.identifier.cristin1332959


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