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Nat Commun. 2018 Oct 2;9(1):4042. doi: 10.1038/s41467-018-06531-4.

Computational design of chemogenetic and optogenetic split proteins.

Author information

1
Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA.
2
Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA.
3
Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA.
4
Department of Neurobiology, Harvard Medical School, Boston, MA, 02115, USA.
5
Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, CA, 94305, USA.
6
Department of Chemistry, University of Pittsburgh, Pittsburgh, PA, 15260, USA.
7
Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA. khahn@med.unc.edu.
8
Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA. khahn@med.unc.edu.
9
Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA. dokh@psu.edu.
10
Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA. dokh@psu.edu.
11
Departments of Pharmacology, and Biochemistry & Molecular Biology, Penn State College of Medicine, Hershey, PA, 17033-0850, USA. dokh@psu.edu.

Abstract

Controlling protein activity with chemogenetics and optogenetics has proven to be powerful for testing hypotheses regarding protein function in rapid biological processes. Controlling proteins by splitting them and then rescuing their activity through inducible reassembly offers great potential to control diverse protein activities. Building split proteins has been difficult due to spontaneous assembly, difficulty in identifying appropriate split sites, and inefficient induction of effective reassembly. Here we present an automated approach to design effective split proteins regulated by a ligand or by light (SPELL). We develop a scoring function together with an engineered domain to enable reassembly of protein halves with high efficiency and with reduced spontaneous assembly. We demonstrate SPELL by applying it to proteins of various shapes and sizes in living cells. The SPELL server (spell.dokhlab.org) offers an automated prediction of split sites.

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PMID:
30279442
PMCID:
PMC6168510
DOI:
10.1038/s41467-018-06531-4
[Indexed for MEDLINE]
Free PMC Article

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