ArticlePloS one2011
Frog oocytes to unveil the structure and supramolecular organization of human transport proteins.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 29 citations in OpenAlex.
- Step-by-Step Protocol for Fluorescence-Based Analysis of Uptake in Transporter-ExpressingBio-protocol · 2026Article
- A pan-vertebrate signaling motif controls the molecular function of intracellular AQP12.The Journal of cell biology · 2026Article
- Real-time tilting and twisting motions of ligand-bound states of α7 nicotinic acetylcholine receptor.European biophysics journal : EBJ · 2024Article
- Mechanistic basis for multidrug resistance and collateral drug sensitivity conferred to the malaria parasite by polymorphisms in PfMDR1 and PfCRT.PLoS biology · 2022Article
- Electrophysiological characterization of a diverse group of sugar transporters from Trichoderma reesei.Scientific reports · 2021Article
- Channelrhodopsin C1C2: Photocycle kinetics and interactions near the central gate.Biophysical journal · 2021Article
- Naphthylphthalamic acid associates with and inhibits PIN auxin transporters.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- Thermophoretic analysis of ligand-specific conformational states of the inhibitory glycine receptor embedded in copolymer nanodiscs.Scientific reports · 2020Article
- Structure of the Regulatory Cytosolic Domain of a Eukaryotic Potassium-Chloride Cotransporter.Structure (London, England : 1993) · 2020Article
- Integrated K+ channel and K+Cl- cotransporter functions are required for the coordination of size and proportion during development.Developmental biology · 2019Article
- Physiological roles and molecular mechanisms of KThe Journal of physiology · 2019Review
- Diverse mutational pathways converge on saturable chloroquine transport via the malaria parasite's chloroquine resistance transporter.Proceedings of the National Academy of Sciences of the United States of America · 2014Article
- Expression, purification, and structural insights for the human uric acid transporter, GLUT9, using the Xenopus laevis oocytes system.PloS one · 2014Article
- Expression, purification and low-resolution structure of human vitamin C transporter SVCT1 (SLC23A1).PloS one · 2013Article
- The biogenesis of potassium transporters: implications of disease-associated mutations.Critical reviews in biochemistry and molecular biologyReview
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Authors and funding
8 authors at 3 institutions in 2 countries.
Funding
Abstract
Structural analyses of heterologously expressed mammalian membrane proteins remain a great challenge given that microgram to milligram amounts of correctly folded and highly purified proteins are required. Here, we present a novel method for the expression and affinity purification of recombinant mammalian and in particular human transport proteins in Xenopus laevis frog oocytes. The method was validated for four human and one murine transporter. Negative stain transmission electron microscopy (TEM) and single particle analysis (SPA) of two of these transporters, i.e., the potassium-chloride cotransporter 4 (KCC4) and the aquaporin-1 (AQP1) water channel, revealed the expected quaternary structures within homogeneous preparations, and thus correct protein folding and assembly. This is the first time a cation-chloride cotransporter (SLC12) family member is isolated, and its shape, dimensions, low-resolution structure and oligomeric state determined by TEM, i.e., by a direct method. Finally, we were able to grow 2D crystals of human AQP1. The ability of AQP1 to crystallize was a strong indicator for the structural integrity of the purified recombinant protein. This approach will open the way for the structure determination of many human membrane transporters taking full advantage of the Xenopus laevis oocyte expression system that generally yields robust functional expression.
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