Artificial Avidin-Based Receptors for a Panel of Small Molecules

Soili I. Lehtonen, Antti Tullila, Nitin Agrawal, Sampo Kukkurainen, Niklas Kähkönen, Masi Koskinen, Tarja K. Nevanen, Mark S Johnson, Tomi Airenne, Markku S. Kulomaa, Tiina A. Riihimäki, Vesa P. Hytönen

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7 Citeringar (Scopus)

Sammanfattning

Proteins with high specificity, affinity, and stability are needed for biomolecular recognition in a plethora of applications. Antibodies are powerful affinity tools, but they may also suffer from limitations such as low stability and high production costs. Avidin and streptavidin provide a promising scaffold for protein engineering, and due to their ultratight binding to D-biotin they are widely used in various biotechnological and biomedical applications. In this study, we demonstrate that the avidin scaffold is suitable for use as a novel receptor for several biologically active small molecules: Artificial, chicken avidin-based proteins, antidins, were generated using a directed evolution method for progesterone, hydrocortisone, testosterone, cholic acid, ketoprofen, and folic acid, all with micromolar to nanomolar affinity and significantly reduced biotin-binding affinity. We also describe the crystal structure of an antidin, sbAvd-2(I117Y), a steroid-binding avidin, which proves that the avidin scaffold can tolerate significant modifications without losing its characteristic tetrameric beta-barrel structure, helping us to further design avidin-based small molecule receptors.
OriginalspråkOdefinierat/okänt
Sidor (från-till)211–221
TidskriftACS Chemical Biology
Volym11
Utgåva1
DOI
StatusPublicerad - 2015
MoE-publikationstypA1 Tidskriftsartikel-refererad

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