Bifunctional avidin with covalently modifiable ligand binding site

Jenni Leppiniemi, Juha A.E. Määttä, Henrik Hammaren, Mikko Soikkeli, Mikko Laitaoja, Janne Jänis, Markku S. Kulomaa, Vesa P. Hytönen

Tutkimustuotos: ArtikkeliScientificvertaisarvioitu

13 Sitaatiot (Scopus)

Abstrakti

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.

AlkuperäiskieliEnglanti
Artikkelie16576
Sivute16576
JulkaisuPLoS One
Vuosikerta6
Numero1
DOI - pysyväislinkit
TilaJulkaistu - 2011
OKM-julkaisutyyppiA1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä

Julkaisufoorumi-taso

  • Ei tasoa

Sormenjälki

Sukella tutkimusaiheisiin 'Bifunctional avidin with covalently modifiable ligand binding site'. Ne muodostavat yhdessä ainutlaatuisen sormenjäljen.

Siteeraa tätä