Modification of the loops in the ligand-binding site turns avidin into a steroid-binding protein


Näytä suppeat kuvailutiedot Riihimäki, Tiina A - Hiltunen, Soili - Rangl, Martina - Nordlund, Henri R - Määttä, Juha A E - Ebner, Andreas - Hinterdorfer, Peter - Kulomaa, Markku S - Takkinen, Kristiina - Hytönen, Vesa P - 2012-06-17T20:14:20Z 2012-06-15 21:55:45 - 2012-06-17T20:14:20Z 2011 -
dc.identifier.issn 1472-6750 -
dc.description BioMed Central Open access -
dc.description.abstract Background Engineered proteins, with non-immunoglobulin scaffolds, have become an important alternative to antibodies in many biotechnical and therapeutic applications. When compared to antibodies, tailored proteins may provide advantageous properties such as a smaller size or a more stable structure. Results Avidin is a widely used protein in biomedicine and biotechnology. To tailor the binding properties of avidin, we have designed a sequence-randomized avidin library with mutagenesis focused at the loop area of the binding site. Selection from the generated library led to the isolation of a steroid-binding avidin mutant (sbAvd-1) showing micromolar affinity towards testosterone (Kd ~ 9 μM). Furthermore, a gene library based on the sbAvd-1 gene was created by randomizing the loop area between β-strands 3 and 4. Phage display selection from this library led to the isolation of a steroid-binding protein with significantly decreased biotin binding affinity compared to sbAvd-1. Importantly, differential scanning calorimetry and analytical gel-filtration revealed that the high stability and the tetrameric structure were preserved in these engineered avidins. Conclusions The high stability and structural properties of avidin make it an attractive molecule for the engineering of novel receptors. This methodology may allow the use of avidin as a universal scaffold in the development of novel receptors for small molecules.   -
dc.language.iso en -
dc.title Modification of the loops in the ligand-binding site turns avidin into a steroid-binding protein -
dc.type fi=Artikkeli aikakauslehdessä | en=Journal article| -
dc.identifier.urn urn:nbn:uta-3-657 -
dc.identifier.doi 10.1186/1472-6750-11-64 -
dc.type.version fi=Kustantajan versio | en=Publisher's version| -
dc.subject.okm fi=Lääketieteen bioteknologia | en=Medical biotechnology| -
dc.administrativeunit fi=Biolääketieteellisen teknologian yksikkö | en=Institute of Biomedical Technology| -
dc.journal.title BMC Biotechnology -
dc.journal.volume 11 -
dc.journal.number 64 -
dc.journal.volumepagerange 1-11 -
dc.oldstats 72 -

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