ArticleScientific reports2013
Function of Shaker potassium channels produced by cell-free translation upon injection into Xenopus oocytes.
Article in Scientific reports, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 22 citations in OpenAlex.
- Evaluation of Cell-Free Synthesized Human Channel Proteins for In Vitro Channel Research.Membranes · 2022Article
- Easy Synthesis of Complex Biomolecular Assemblies: Wheat Germ Cell-Free Protein Expression in Structural Biology.Frontiers in molecular biosciences · 2021Review
- Article
- A heterodimeric glutathioneThe Journal of biological chemistry · 2019Article
- Purification and Reconstitution of TRPV1 for Spectroscopic Analysis.Journal of visualized experiments : JoVE · 2018Article
- LRET Determination of Molecular Distances during pH Gating of the Mammalian Inward Rectifier Kir1.1b.Biophysical journal · 2018Article
- Whole-GUV patch-clamping.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- Shock Wave-Induced Damage of a Protein by Void Collapse.Biophysical journal · 2016Article
- Cell-free expression of a functional pore-only sodium channel.Protein expression and purification · 2015Article
- Production of monoclonal antibodies against GPCR using cell-free synthesized GPCR antigen and biotinylated liposome-based interaction assay.Scientific reports · 2015Article
- Expression platforms for producing eukaryotic proteins: a comparison of E. coli cell-based and wheat germ cell-free synthesis, affinity and solubility tags, and cloning strategies.Journal of structural and functional genomics · 2015Article
- Direct injection of cell-free Kir1.1 protein into Xenopus oocytes replicates single-channel currents derived from Kir1.1 mRNA.Channels (Austin, Tex.) · 2015Article
- Site-directed spin labeling reveals pentameric ligand-gated ion channel gating motions.PLoS biology · 2013Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
Voltage-gated ion channels are a class of membrane proteins that temporally orchestrate the ion flux critical for chemical and electrical signaling in excitable cells. Current methods to investigate the function of these channels rely on heterologous expression in living systems or reconstitution into artificial membranes; however these approaches have inherent drawbacks which limit potential biophysical applications. Here, we describe a new integrated approach combining cell-free translation of membrane proteins and in vivo expression using Xenopus laevis oocytes. In this method, proteoliposomes containing Shaker potassium channels are synthesized in vitro and injected into the oocytes, yielding functional preparations as shown by electrophysiological and fluorescence measurements within few hours. This strategy for studying eukaryotic ion channels is contrasted with existing, laborious procedures that require membrane protein extraction and reconstitution into synthetic lipid systems.
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