Abstract
AbstractMonobodies 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.
Funder
U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences
NSF | ENG/OAD | Division of Chemical, Bioengineering, Environmental, and Transport Systems
DOE | SC | Biological and Environmental Research
The Pew Charitable Trusts, The Eric and Wendy Schmidt Transformative Technology Fund Award, and the Camille Dreyfus Teacher-Scholar Award
Publisher
Springer Science and Business Media LLC
Subject
General Physics and Astronomy,General Biochemistry, Genetics and Molecular Biology,General Chemistry
Cited by
43 articles.
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