TY - JOUR
T1 - Engineering extrinsic disorder to control protein activity in living cells
AU - Dagliyan, Onur
AU - Tarnawski, Miroslaw
AU - Chu, Pei Hsuan
AU - Shirvanyants, David
AU - Schlichting, Ilme
AU - Dokholyan, Nikolay V.
AU - Hahn, Klaus M.
N1 - Funding Information:
This work was supported by NIH grants P01-GM103723 and P41-EB002025 (K.M.H.), R01GM080742 (N.V.D.), and by Deutsche Forschungsgemeinschaft grant FOR1279 (I.S.). O.D. is a Howard Hughes Medical Institute International Student Research Fellow. X-ray data were collected at the Swiss Light Source, beamline X10SA and X12SA, Paul Scherrer Institute, Villigen, Switzerland. We thank A. Menzel and T. Barends for collecting the small angle x-ray scattering data, and M. Shobair for his help with PI-Vav2 simulations. Atomic coordinates and structure factors have been deposited in the Protein Data Bank under accession codes: 5HZJ (PI-ITSN1-WT), 5HZI (PI-ITSN1-C450M), 5HZK (PI-ITSN1-WT: Cdc42), and 5HZH (PI-Rac1-C450A). O.D., N.V.D., and K.M.H. designed the research; M.T. performed the crystallographic work and conducted hydrogen-deuterium exchange coupled to mass spectrometry assays under the directions of I.S. who collected the x-ray data; P.-H.C. helped the characterization of PI-Src; D.S. conducted the simulations of PI-Src; O.D. performed all experiments and computations not listed above; O.D. and K.M.H. wrote the manuscript with input from all authors. The constructs described in this paper are available from K.M.H under a material transfer agreement with the University of North Carolina-Chapel Hill.
Publisher Copyright:
Copyright 2016 by the American Association for the Advancement of Science; all rights reserved.
PY - 2016/12/16
Y1 - 2016/12/16
N2 - 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 asmodulated themorphodynamics of living cells.Our results illustrate a broadly applicable approach to design physiological protein switches.
AB - 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 asmodulated themorphodynamics of living cells.Our results illustrate a broadly applicable approach to design physiological protein switches.
UR - http://www.scopus.com/inward/record.url?scp=85006320532&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85006320532&partnerID=8YFLogxK
U2 - 10.1126/science.aah3404
DO - 10.1126/science.aah3404
M3 - Article
C2 - 27980211
AN - SCOPUS:85006320532
SN - 0036-8075
VL - 354
SP - 1441
EP - 1444
JO - Science
JF - Science
IS - 6318
ER -