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

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,...

Full description

Saved in:
Bibliographic Details
Published in:PloS one 2019-02, Vol.14 (2), p.e0212339-e0212339
Main Authors: Agrawal, Nitin, Lehtonen, Soili I, Uusi-Mäkelä, Meri, Jain, Purvi, Viitala, Sari, Määttä, Juha A E, Kähkönen, Niklas, Azizi, Latifeh, Riihimäki, Tiina A, Kulomaa, Markku S, Johnson, Mark S, Hytönen, Vesa P, Airenne, Tomi T
Format: Article
Language:English
Subjects:
Affinity
Analysis
Animals
Avidin
Avidin - chemistry
Avidin - metabolism
Binding
Binding proteins
Binding Sites
Biochemistry
Bioinformatics
Biological Assay
Biology and Life Sciences
Biotin
Biotin - chemistry
Biotin - metabolism
Chickens
Computer applications
Crystallization
Crystallography
Dogs
Laboratories
Ligands
Medicine
Models, Molecular
Molecular biology
Mutagenesis
Phage display
Phages
Pharmaceutical sciences
Physical Sciences
Polypeptides
Poultry
Progesterone
Progesterone - blood
Progesterone - chemistry
Progesterone - metabolism
Protein Binding
Proteins
Research and Analysis Methods
Sex hormones
Steroid hormones
Steroids
Streptavidin
Structure
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0212339