Bifunctional Avidin with Covalently Modifiable Ligand Binding Site


Näytä suppeat kuvailutiedot Leppiniemi, Jenni - Määttä, Juha A E - Hammaren, Henrik - Soikkeli, Mikko - Laitaoja, Mikko - Jänis, Janne - Kulomaa, Markku S - Hytönen, Vesa P - 2012-06-17T20:14:34Z 2012-06-15 03:38:38 - 2012-06-17T20:14:34Z 2011 -
dc.identifier.issn 1932-6203 -
dc.description Public Library of Science -
dc.description.abstract The extensive use of avidin and streptavidin in life sciences originates from the extraordinary tight biotin-binding affinity of these tetrameric proteins. Numerous studies have been performed to modify the biotin-binding affinity of (strept)avidin to improve the existing applications. Even so, (strept)avidin greatly favours its natural ligand, biotin. Here we engineered the biotin-binding pocket of avidin with a single point mutation S16C and thus introduced a chemically active thiol group, which could be covalently coupled with thiol-reactive molecules. This approach was applied to the previously reported bivalent dual chain avidin by modifying one binding site while preserving the other one intact. Maleimide was then coupled to the modified binding site resulting in a decrease in biotin affinity. Furthermore, we showed that this thiol could be covalently coupled to other maleimide derivatives, for instance fluorescent labels, allowing intratetrameric FRET. The bifunctional avidins described here provide improved and novel tools for applications such as the biofunctionalization of surfaces. -
dc.language.iso en -
dc.title Bifunctional Avidin with Covalently Modifiable Ligand Binding Site -
dc.type fi=Artikkeli aikakauslehdessä | en=Journal article| -
dc.identifier.urn urn:nbn:uta-3-702 -
dc.identifier.doi 10.1371/journal.pone.0016576 -
dc.type.version fi=Kustantajan versio | en=Publisher's version| -
dc.subject.okm fi=Biokemia, solu- ja molekyylibiologia | en=Biochemistry, cell and molecular biology| -
dc.administrativeunit fi=Biolääketieteellisen teknologian yksikkö | en=Institute of Biomedical Technology| -
dc.journal.title PLoS ONE -
dc.journal.volume 6 -
dc.journal.number 1 -
dc.journal.volumepagerange 1-11 -
dc.oldstats 51 -

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