Modular transient nanoclustering of activated β2-adrenergic receptors revealed by single-molecule tracking of conformation-specific nanobodies
Copyright policy )Modular transient nanoclustering of activated β2-adrenergic receptors revealed by single-molecule tracking of conformation-specific nanobodies
SignificanceProteins moving freely on the plasma membrane can become transiently trapped in functionally essential clusters. This capability is likely to be influenced by subtle conformational states of the protein promoting or preventing such confinement. The downside of conventional imaging of overexpressed tagged proteins is that it precludes selective tracking of inherently minor albeit functionally essential conformer populations. Intracellular expression of single-chain nanobodies allowed us to track endogenous proteins in highly specific conformational states in live cells and small organisms. We unveiled the full scope of nanoclustering behavior of β2-adrenergic receptors in various conformations, along with their transient nature. This technique is broadly applicable to other proteins and will help unravel essential dynamics and organization of nanoclusters.
Models, Molecular, Protein Conformation, Recombinant Fusion Proteins, Fluorescent Antibody Technique, Gene Expression, Cell Line, single-particle−tracking superresolution microscopy, Mice, Genes, Reporter, Animals, Humans, β2-adrenoreceptor, Zebrafish, Science & Technology, Membrane Proteins, Nanobiotechnology, Single-Domain Antibodies, single-particle-tracking superresolution microscopy, nanobodies, Endocytosis, Single Molecule Imaging, Multidisciplinary Sciences, 1000 General, Science & Technology - Other Topics, Receptors, Adrenergic, beta-2, Protein Binding
Models, Molecular, Protein Conformation, Recombinant Fusion Proteins, Fluorescent Antibody Technique, Gene Expression, Cell Line, single-particle−tracking superresolution microscopy, Mice, Genes, Reporter, Animals, Humans, β2-adrenoreceptor, Zebrafish, Science & Technology, Membrane Proteins, Nanobiotechnology, Single-Domain Antibodies, single-particle-tracking superresolution microscopy, nanobodies, Endocytosis, Single Molecule Imaging, Multidisciplinary Sciences, 1000 General, Science & Technology - Other Topics, Receptors, Adrenergic, beta-2, Protein Binding
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