dc.contributor.author | Schrader, S | |
dc.contributor.author | Tiralongo, E | |
dc.contributor.author | Paris, G | |
dc.contributor.author | Yoshino, T | |
dc.contributor.author | Schauer, R | |
dc.date.accessioned | 2017-05-03T14:35:26Z | |
dc.date.available | 2017-05-03T14:35:26Z | |
dc.date.issued | 2003 | |
dc.identifier.issn | 0003-2697 | |
dc.identifier.doi | 10.1016/j.ab.2003.07.016 | |
dc.identifier.uri | http://hdl.handle.net/10072/24068 | |
dc.description.abstract | Trans-sialidase (E.C. 3.2.1.18) catalyzes the transfer of preferably a2,3-linked sialic acid to another glycan or glycoconjugate, forming a new a2,3 linkage to galactose or N-acetylgalactosamine. Here, we describe a nonradioactive 96-well plate fluorescence test for monitoring trans-sialidase activity with high sensitivity, specificity, and reproducibility using sialyllactose and 4-methylumbelliferyl-߭Image -galactoside as donor and acceptor substrates, respectively. The assay conditions were optimized using the trans-sialidase from Trypanosoma congolense and its general applicability was confirmed with recombinant trans-sialidase from Trypanosoma cruzi. Using this procedure, a large number of samples can be tested quickly and reliably, for instance in monitoring trans-sialidase during enzyme purification and the production of monoclonal antibodies, for enzyme characterization, and for identifying potential substrates and inhibitors. The trans-sialidase assay reported here was capable of detecting trans-sialidase activity in the low-mU range and may be a valuable tool in the search for further trans-sialidases in various biological systems. | |
dc.description.peerreviewed | Yes | |
dc.description.publicationstatus | Yes | |
dc.language | English | |
dc.language.iso | eng | |
dc.publisher | Academic Press | |
dc.publisher.place | USA | |
dc.relation.ispartofpagefrom | 139 | |
dc.relation.ispartofpageto | 147 | |
dc.relation.ispartofjournal | Analytical Biochemistry | |
dc.relation.ispartofvolume | 322 | |
dc.subject.fieldofresearch | Analytical chemistry | |
dc.subject.fieldofresearch | Other chemical sciences | |
dc.subject.fieldofresearch | Biochemistry and cell biology | |
dc.subject.fieldofresearchcode | 3401 | |
dc.subject.fieldofresearchcode | 3499 | |
dc.subject.fieldofresearchcode | 3101 | |
dc.title | A nonradioactive 96-well plate assay for screening of trans-sialidase activity | |
dc.type | Journal article | |
dc.type.description | C1 - Articles | |
dc.type.code | C - Journal Articles | |
gro.date.issued | 2015-02-06T01:36:46Z | |
gro.hasfulltext | No Full Text | |
gro.griffith.author | Tiralongo, Evelin | |