Development of light-responsive protein binding in the monobody non-immunoglobulin scaffold.

Publication Year
2020

Type

Journal Article
Abstract

Monobodies are synthetic non-immunoglobulin customizable protein binders invaluable to basic and applied research, and of considerable potential as future therapeutics and diagnostic tools. The ability to reversibly control their binding activity to their targets on demand would significantly expand their applications in biotechnology, medicine, and research. Here we present, as proof-of-principle, the development of a light-controlled monobody (OptoMB) that works in vitro and in cells and whose affinity for its SH2-domain target exhibits a 330-fold shift in binding affinity upon illumination. We demonstrate that our αSH2-OptoMB can be used to purify SH2-tagged proteins directly from crude E. coli extract, achieving 99.8% purity and over 40% yield in a single purification step. By virtue of their ability to be designed to bind any protein of interest, OptoMBs have the potential to find new powerful applications as light-switchable binders of untagged proteins with the temporal and spatial precision afforded by light.

Journal
Nat Commun
Volume
11
Issue
1
Pages
4045
Date Published
2020 Aug 13
ISSN Number
2041-1723
Alternate Journal
Nat Commun
PMCID
PMC7427095
PMID
32792484