title: Engineering extrinsic disorder to control protein activity in living cells.
abstract: Optogenetic and chemogenetic control of proteins has revealed otherwise inaccessible facets of signaling dynamics. Here, we use light- or ligand-sensitive domains to modulate the structural disorder of diverse proteins, thereby generating robust allosteric switches. Sensory domains were inserted into nonconserved, surface-exposed loops that were tight and identified computationally as allosterically coupled to active sites. Allosteric switches introduced into motility signaling proteins (kinases, guanosine triphosphatases, and guanine exchange factors) controlled conversion between conformations closely resembling natural active and inactive states, as well as modulated the morphodynamics of living cells. Our results illustrate a broadly applicable approach to design physiological protein switches.
authors.affiliation: Program in Molecular and Cellular Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.|Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.|Department of Pharmacology and Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
authors.forename: Onur
authors.initials: O
authors.lastname: Dagliyan
authors.affiliation: Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Jahnstrasse 29, 69120 Heidelberg, Germany.
authors.forename: Miroslaw
authors.initials: M
authors.lastname: Tarnawski
authors.affiliation: Department of Pharmacology and Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
authors.forename: Pei-Hsuan
authors.initials: PH
authors.lastname: Chu
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
authors.forename: David
authors.initials: D
authors.lastname: Shirvanyants
authors.affiliation: Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Jahnstrasse 29, 69120 Heidelberg, Germany.
authors.forename: Ilme
authors.initials: I
authors.lastname: Schlichting
authors.affiliation: Program in Molecular and Cellular Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA. khahn@med.unc.edu dokh@email.unc.edu.|Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
authors.forename: Nikolay V
authors.initials: NV
authors.lastname: Dokholyan
authors.affiliation: Department of Pharmacology and Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA. khahn@med.unc.edu dokh@email.unc.edu.
authors.forename: Klaus M
authors.initials: KM
authors.lastname: Hahn
mesh_terms: D000494:Allosteric Regulation / Q000235:genetics / Q000528:radiation effects; D000495:Allosteric Site; D020134:Catalytic Domain; D004789:Enzyme Activation / Q000235:genetics / Q000528:radiation effects; D020558:GTP Phosphohydrolases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000528:radiation effects; D020662:Guanine Nucleotide Exchange Factors / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D008024:Ligands; D008027:Light; D062308:Optogenetics; D000072417:Protein Domains / Q000528:radiation effects; D015202:Protein Engineering; D051567:Proto-Oncogene Proteins c-vav / Q000737:chemistry; D015398:Signal Transduction; D019061:src-Family Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000528:radiation effects
chemical_list: D020662:Guanine Nucleotide Exchange Factors; D008024:Ligands; D051567:Proto-Oncogene Proteins c-vav; C093651:VAV2 protein, human; D019061:src-Family Kinases; D020558:GTP Phosphohydrolases