Molecular features of steroid-binding antidins and their use for assaying serum progesterone

Nitin Agrawal, SI Lehtonen, M Uusi-Mäkelä, P Jain, S Viitala, Määttä JAE, N Kähkönen, L Azizi, TA Riihimäki, MS Kulomaa, Mark S Johnson, VP Hytönen, Tomi Airenne

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Abstract

Chicken avidin (Avd) and streptavidin from Streptomyces avidinii are extensively used in bionanotechnology due to their extremely tight binding to biotin (Kd ~ 10-15 M for chicken Avd). We previously reported engineered Avds known as antidins, which have micro- to nanomolar affinities for steroids, non-natural ligands of Avd. Here, we report the 2.8 Å X-ray structure of the sbAvd-2 (I117Y) antidin co-crystallized with progesterone. We describe the creation of new synthetic phage display libraries and report the experimental as well as computational binding analysis of progesterone-binding antidins. We introduce a next-generation antidin with 5 nM binding affinity for progesterone, and demonstrate the use of antidins for measuring progesterone in serum samples. Our data give insights on how to engineer and alter the binding preferences of Avds and to develop better molecular tools for modern bionanotechnological applications.

Original languageUndefined/Unknown
Pages (from-to)
JournalPLoS ONE
Volume14
Issue number2
DOIs
Publication statusPublished - 2019
MoE publication typeA1 Journal article-refereed

Keywords

  • 3D structural analysis
  • ligand binding
  • avidin-biotin technology
  • Diagnostics
  • Biochemistry & Molecular Biology

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