Blog
About

21
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: not found

      Evolving the lock to fit the key to create a family of G protein-coupled receptors potently activated by an inert ligand.

      Proceedings of the National Academy of Sciences of the United States of America

      Animals, Cell Line, Transformed, Clozapine, analogs & derivatives, pharmacology, Designer Drugs, Epitopes, Evolution, Molecular, G Protein-Coupled Inwardly-Rectifying Potassium Channels, metabolism, Hippocampus, drug effects, Humans, Hydrolysis, Ligands, Models, Molecular, Mutant Proteins, Myocytes, Smooth Muscle, Neurons, Phosphatidylinositols, Protein Engineering, Pulmonary Artery, Saccharomyces cerevisiae, cytology, Rats, Receptor, Muscarinic M3, Receptor, Muscarinic M4, Receptors, G-Protein-Coupled

      Read this article at

      ScienceOpenPublisherPMC
      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          We evolved muscarinic receptors in yeast to generate a family of G protein-coupled receptors (GPCRs) that are activated solely by a pharmacologically inert drug-like and bioavailable compound (clozapine-N-oxide). Subsequent screening in human cell lines facilitated the creation of a family of muscarinic acetylcholine GPCRs suitable for in vitro and in situ studies. We subsequently created lines of telomerase-immortalized human pulmonary artery smooth muscle cells stably expressing all five family members and found that each one faithfully recapitulated the signaling phenotype of the parent receptor. We also expressed a G(i)-coupled designer receptor in hippocampal neurons (hM(4)D) and demonstrated its ability to induce membrane hyperpolarization and neuronal silencing. We have thus devised a facile approach for designing families of GPCRs with engineered ligand specificities. Such reverse-engineered GPCRs will prove to be powerful tools for selectively modulating signal-transduction pathways in vitro and in vivo.

          Related collections

          Author and article information

          Journal
          17360345
          1829280
          10.1073/pnas.0700293104

          Comments

          Comment on this article