Construction of Chimeric Dual-Chain Avidin by Tandem Fusion of the Related Avidins


Näytä suppeat kuvailutiedot Riihimäki, Tiina A - Kukkurainen, Sampo - Varjonen, Suvi - Hörhä, Jarno - Nyholm, Thomas KM - Kulomaa, Markku S - Hytönen, Vesa P - 2012-06-17T20:14:37Z 2012-06-15 06:59:56 - 2012-06-17T20:14:37Z 2011 -
dc.identifier.issn 1932-6203 -
dc.description Public Library of Science -
dc.description.abstract Background Avidin is a chicken egg-white protein with high affinity to vitamin H, also known as D-biotin. Many applications in life science research are based on this strong interaction. Avidin is a homotetrameric protein, which promotes its modification to symmetrical entities. Dual-chain avidin, a genetically engineered avidin form, has two circularly permuted chicken avidin monomers that are tandem-fused into one polypeptide chain. This form of avidin enables independent modification of the two domains, including the two biotin-binding pockets; however, decreased yields in protein production, compared to wt avidin, and complicated genetic manipulation of two highly similar DNA sequences in the tandem gene have limited the use of dual-chain avidin in biotechnological applications. Principal Findings To overcome challenges associated with the original dual-chain avidin, we developed chimeric dual-chain avidin, which is a tandem fusion of avidin and avidin-related protein 4 (AVR4), another member of the chicken avidin gene family. We observed an increase in protein production and better thermal stability, compared with the original dual-chain avidin. Additionally, PCR amplification of the hybrid gene was more efficient, thus enabling more convenient and straightforward modification of the dual-chain avidin. When studied closer, the generated chimeric dual-chain avidin showed biphasic biotin dissociation. Significance The improved dual-chain avidin introduced here increases its potential for future applications. This molecule offers a valuable base for developing bi-functional avidin tools for bioseparation, carrier proteins, and nanoscale adapters. Additionally, this strategy could be helpful when generating hetero-oligomers from other oligomeric proteins with high structural similarity. -
dc.language.iso en -
dc.title Construction of Chimeric Dual-Chain Avidin by Tandem Fusion of the Related Avidins -
dc.type fi=Artikkeli aikakauslehdessä | en=Journal article| -
dc.identifier.urn urn:nbn:uta-3-711 -
dc.identifier.doi 10.1371/journal.pone.0020535 -
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 5 -
dc.journal.volumepagerange 1-10 -
dc.oldstats 62 -

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