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research article

Computational design of ligand-binding proteins with high affinity and selectivity

Tinberg, Christine E.
•
Khare, Sagar D.
•
Dou, Jiayi
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2013
Nature

The ability to design proteins with high affinity and selectivity for any given small molecule is a rigorous test of our understanding of the physiochemical principles that govern molecular recognition. Attempts to rationally design ligand-binding proteins have met with little success, however, and the computational design of protein-small-molecule interfaces remains an unsolved problem. Current approaches for designing ligand-binding proteins for medical and biotechnological uses rely on raising antibodies against a target antigen in immunized animals and/or performing laboratory-directed evolution of proteins with an existing low affinity for the desired ligand, neither of which allows complete control over the interactions involved in binding. Here we describe a general computational method for designing pre-organized and shape complementary small-molecule-binding sites, and use it to generate protein binders to the steroid digoxigenin (DIG). Of seventeen experimentally characterized designs, two bind DIG; the model of the higher affinity binder has the most energetically favourable and pre-organized interface in the design set. A comprehensive binding-fitness landscape of this design, generated by library selections and deep sequencing, was used to optimize its binding affinity to a picomolar level, and X-ray co-crystal structures of two variants show atomic-level agreement with the corresponding computational models. The optimized binder is selective for DIG over the related steroids digitoxigenin, progesterone and β-oestradiol, and this steroid binding preference can be reprogrammed by manipulation of explicitly designed hydrogen-bonding interactions. The computational design method presented here should enable the development of a new generation of biosensors, therapeutics and diagnostics.

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Type
research article
DOI
10.1038/nature12443
Web of Science ID

WOS:000324244900039

Author(s)
Tinberg, Christine E.
Khare, Sagar D.
Dou, Jiayi
Doyle, Lindsey
Nelson, Jorgen W.
Schena, Alberto  
Jankowski, Wojciech
Kalodimos, Charalampos G.
Johnsson, Kai  
Stoddard, Barry L.
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Date Issued

2013

Publisher

Nature Publishing Group

Published in
Nature
Volume

501

Issue

7466

Start page

212

End page

6

Editorial or Peer reviewed

NON-REVIEWED

Written at

EPFL

EPFL units
LIP  
Available on Infoscience
September 6, 2013
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/94477
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