TY - JOUR
T1 - Defining and Exploiting Hypersensitivity Hotspots to Facilitate Abscisic Acid Agonist Optimization
AU - Elzinga, Dezi
AU - Sternburg, Erin
AU - Sabbadin, Davide
AU - Bartsch, Michael
AU - Park, Sang Youl
AU - Vaidya, Aditya
AU - Mosquna, Assaf
AU - Kaundal, Amita
AU - Wendeborn, Sebastian
AU - Lachia, Mathilde
AU - Karginov, Fedor V.
AU - Cutler, Sean R.
N1 - Publisher Copyright:
© 2019 American Chemical Society.
PY - 2019/3/15
Y1 - 2019/3/15
N2 - Pyrabactin resistance 1 (PYR1) and related abscisic acid (ABA) receptors are new targets for manipulating plant drought tolerance. Here, we identify and use PYR1 hypersensitive mutants to define ligand binding hotspots and show that these can guide improvements in agonist potency. One hotspot residue defined, A160, is part of a pocket that is occupied by ABA's C6 methyl or by the toluyl methyl of the synthetic agonist quinabactin (QB). A series of QB analogues substituted at the toluyl position were synthesized and provide up to 10-fold gain in activity in vitro. Furthermore, we demonstrate that hypersensitive receptors can be used to improve the sensitivity of a previously described mammalian cell ABA-regulated transcriptional circuit by three orders of magnitude. Collectively, our data show that the systematic mapping of hypersensitivity sites in a ligand-binding pocket can help guide ligand optimization and tune the sensitivity of engineered receptors.
AB - Pyrabactin resistance 1 (PYR1) and related abscisic acid (ABA) receptors are new targets for manipulating plant drought tolerance. Here, we identify and use PYR1 hypersensitive mutants to define ligand binding hotspots and show that these can guide improvements in agonist potency. One hotspot residue defined, A160, is part of a pocket that is occupied by ABA's C6 methyl or by the toluyl methyl of the synthetic agonist quinabactin (QB). A series of QB analogues substituted at the toluyl position were synthesized and provide up to 10-fold gain in activity in vitro. Furthermore, we demonstrate that hypersensitive receptors can be used to improve the sensitivity of a previously described mammalian cell ABA-regulated transcriptional circuit by three orders of magnitude. Collectively, our data show that the systematic mapping of hypersensitivity sites in a ligand-binding pocket can help guide ligand optimization and tune the sensitivity of engineered receptors.
UR - http://www.scopus.com/inward/record.url?scp=85062959467&partnerID=8YFLogxK
U2 - 10.1021/acschembio.8b00955
DO - 10.1021/acschembio.8b00955
M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???
C2 - 30668093
AN - SCOPUS:85062959467
SN - 1554-8929
VL - 14
SP - 332
EP - 336
JO - ACS Chemical Biology
JF - ACS Chemical Biology
IS - 3
ER -