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Nature Chemical Biology Jan 2024Plants sense abscisic acid (ABA) using chemical-induced dimerization (CID) modules, including the receptor PYR1 and HAB1, a phosphatase inhibited by ligand-activated...
Plants sense abscisic acid (ABA) using chemical-induced dimerization (CID) modules, including the receptor PYR1 and HAB1, a phosphatase inhibited by ligand-activated PYR1. This system is unique because of the relative ease with which ligand recognition can be reprogrammed. To expand the PYR1 system, we designed an orthogonal '*' module, which harbors a dimer interface salt bridge; X-ray crystallographic, biochemical and in vivo analyses confirm its orthogonality. We used this module to create PYR1*/HAB1* and PYR1*/HAB1*, which possess nanomolar sensitivities to their activating ligands mandipropamid and azinphos-ethyl. Experiments in Arabidopsis thaliana and Saccharomyces cerevisiae demonstrate the sensitive detection of banned organophosphate contaminants using living biosensors and the construction of multi-input/output genetic circuits. Our new modules enable ligand-programmable multi-channel CID systems for plant and eukaryotic synthetic biology that can empower new plant-based and microbe-based sensing modalities.
Topics: Abscisic Acid; Arabidopsis; Arabidopsis Proteins; Dimerization; Ligands; Membrane Transport Proteins
PubMed: 37872402
DOI: 10.1038/s41589-023-01447-7 -
Toxicology and Applied Pharmacology Sep 2001Ethylazinphos increases the passive proton permeability of lipid bilayers reconstituted with dipalmitoylphosphatidylcholine (DPPC) and mitochondrial lipids. A sharp...
Ethylazinphos increases the passive proton permeability of lipid bilayers reconstituted with dipalmitoylphosphatidylcholine (DPPC) and mitochondrial lipids. A sharp increase of proton permeability is detected at insecticide/lipid molar ratios identical to those inducing phase separation in the plane of DPPC bilayers, as revealed by differential scanning calorimetry (DSC). Ethylazinphos progressively depresses the transmembrane potential (DeltaPsi) of mitochondria supported by piruvate/malate, succinate, or ascorbate/TMPD. Additionally, a decreased depolarization induced by ADP depends on ethylazinphos concentration, reflecting a phosphorylation depression. This loss of phosphorylation is a consequence of a decreased DeltaPsi. A decreased respiratory control ratio is also observed, since ethylazinphos stimulates state 4 respiration and inhibits ADP-stimulated respiration (state 3). Ethylazinphos concentrations up to 100 nmol/mg mitochondrial protein increase the rate of state 4 together with a decrease in DeltaPsi, without significant perturbation of state 3 and carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP)-uncoupled respiration. For increased insecticide concentrations, the state 3 and FCCP-uncoupled respiration are inhibited to approximately the same extent. The perturbations are more pronounced when the energization is supported by pyruvate/malate and less effective when succinate is used as substrate. The present data, in association with previous DSC studies, indicate that ethylazinphos, at concentrations up to 100 nmol/mg mitochondrial protein, interacts with the lipid bilayer of mitochondrial membrane, changing the lipid organization and increasing the proton permeability of the inner membrane. The increased proton permeability explains the decreased oxidative phosphorylation coupling. Resulting disturbed ATP synthesis may significantly underlie the mechanisms of ethylazinphos toxicity, since most of cell energy in eukaryotes is provided by mitochondria.
Topics: Animals; Azinphosmethyl; Calorimetry, Differential Scanning; Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; Cytoplasmic Vesicles; Dose-Response Relationship, Drug; Female; Insecticides; Intracellular Membranes; Male; Membrane Lipids; Membrane Potentials; Mitochondria, Liver; Oxidative Phosphorylation; Permeability; Protons; Rats; Rats, Sprague-Dawley
PubMed: 11559019
DOI: 10.1006/taap.2001.9246 -
Biochimica Et Biophysica Acta May 1996The interaction of ethylazinphos with the physical organization of model and native membranes was investigated by means of fluorescence polarization of...
The interaction of ethylazinphos with the physical organization of model and native membranes was investigated by means of fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) and of its propionic acid derivative (DPH-PA). Ethylazinphos shifts the phase transition midpoint to lower temperature values and broadens the phase transition profile of bilayers reconstituted with dimyristoyl-, dipalmitoyl- and distearoylphosphatidylcholines (DMPC, DPPC, DSPC), as detected by DPH and DPH-PA. Additionally, both probes detect significant effects of ethylazinphos in the fluid phase of the above lipid bilayers. The insecticide perturbations are more pronounced in bilayers of short-chain lipids, e.g., DMPC, in correlation with the higher partition in these membranes. On the other hand, the insecticide increases to some extent the ordering promoted by cholesterol in the fluid phase of DMPC, but high cholesterol concentrations (> or = 30 mol%) almost prevent insecticide interaction, as revealed by DPH and DPH-PA. In agreement with the results in models of synthetic lipids, the increase of intrinsic cholesterol in fluid native membranes depresses the partition values of ethylazinphos and consequently its effects.
Topics: Azinphosmethyl; Cell Membrane; Cholesterol; Diphenylhexatriene; Fluorescence Polarization; Fluorescent Dyes; Insecticides; Lipid Bilayers; Phospholipids
PubMed: 8652607
DOI: 10.1016/0005-2736(96)00012-0