One-step nanoscale expansion microscopy reveals individual protein shapes.

Autor:	Shaib AH; Institute for Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany. ali.shaib@med.uni-goettingen.de., Chouaib AA; Department of Cellular Neurophysiology, Center for Integrative Physiology and Molecular Medicine (CIPMM), Saarland University, Homburg, Germany., Chowdhury R; Institute for Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany.; Department of Chemistry, GITAM School of Science, GITAM, Hyderabad, India., Altendorf J; Institute for Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany., Mihaylov D; MRC Laboratory of Molecular Biology, Cambridge, UK., Zhang C; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA, USA.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.; McGovern Institute, Massachusetts Institute of Technology, Cambridge, MA, USA.; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA, USA.; Koch Institute, Massachusetts Institute of Technology, Cambridge, MA, USA.; Center for Neurobiological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA., Krah D; Institute for Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany., Imani V; Institute for Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany., Spencer RKW; Institute for Theoretical Physics, Georg-August University, Göttingen, Germany., Georgiev SV; Institute for Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany., Mougios N; Institute for Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany.; Center for Biostructural Imaging of Neurodegeneration, University Medical Center Göttingen, Göttingen, Germany., Monga M; Biochemistry of Membrane Dynamics Group, Institute for Auditory Neuroscience, University Medical Center Göttingen, Göttingen, Germany., Reshetniak S; Institute for Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany., Mimoso T; Institute for X-Ray Physics, University of Göttingen, Göttingen, Germany., Chen H; Institute for Auditory Neuroscience and InnerEarLab, University Medical Center Göttingen, Göttingen, Germany., Fatehbasharzad P; Institute for Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany., Crzan D; Institute for Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany., Saal KA; Institute for Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany., Alawieh MM; Institute for Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany., Alawar N; Department of Cellular Neurophysiology, Center for Integrative Physiology and Molecular Medicine (CIPMM), Saarland University, Homburg, Germany., Eilts J; Department of Biotechnology and Biophysics, Biocenter, University of Würzburg, Am Hubland, Würzburg, Germany., Kang J; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA, USA.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.; McGovern Institute, Massachusetts Institute of Technology, Cambridge, MA, USA.; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA, USA.; Koch Institute, Massachusetts Institute of Technology, Cambridge, MA, USA.; Center for Neurobiological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA., Soleimani A; Institute for Theoretical Physics, Georg-August University, Göttingen, Germany., Müller M; Institute for Theoretical Physics, Georg-August University, Göttingen, Germany., Pape C; Institute of Computer Science, Georg-August University Göttingen, Göttingen, Germany., Alvarez L; Leica Microsystems CMS GmbH, Mannheim, Germany., Trenkwalder C; Department of Neurosurgery, University Medical Center, Göttingen, Germany.; Paracelsus-Elena-Klinik, Kassel, Germany., Mollenhauer B; Paracelsus-Elena-Klinik, Kassel, Germany.; Department of Neurology, University Medical Center, Göttingen, Germany., Outeiro TF; Department of Experimental Neurodegeneration, Center for Biostructural Imaging of Neurodegeneration, University Medical Center Göttingen, Göttingen, Germany.; Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany.; Translational and Clinical Research Institute, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, UK.; German Center for Neurodegenerative Diseases (DZNE), Göttingen, Germany., Köster S; Institute for X-Ray Physics, University of Göttingen, Göttingen, Germany.; Cluster of Excellence 'Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells' (MBExC), University of Göttingen, Göttingen, Germany., Preobraschenski J; Biochemistry of Membrane Dynamics Group, Institute for Auditory Neuroscience, University Medical Center Göttingen, Göttingen, Germany.; Cluster of Excellence 'Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells' (MBExC), University of Göttingen, Göttingen, Germany., Becherer U; Department of Cellular Neurophysiology, Center for Integrative Physiology and Molecular Medicine (CIPMM), Saarland University, Homburg, Germany., Moser T; Biochemistry of Membrane Dynamics Group, Institute for Auditory Neuroscience, University Medical Center Göttingen, Göttingen, Germany.; Cluster of Excellence 'Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells' (MBExC), University of Göttingen, Göttingen, Germany.; Auditory Neuroscience and Synaptic Nanophysiology Group, Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany., Boyden ES; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA, USA.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.; McGovern Institute, Massachusetts Institute of Technology, Cambridge, MA, USA.; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA, USA.; Koch Institute, Massachusetts Institute of Technology, Cambridge, MA, USA.; Center for Neurobiological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA., Aricescu AR; MRC Laboratory of Molecular Biology, Cambridge, UK., Sauer M; Department of Biotechnology and Biophysics, Biocenter, University of Würzburg, Am Hubland, Würzburg, Germany., Opazo F; Institute for Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany.; Center for Biostructural Imaging of Neurodegeneration, University Medical Center Göttingen, Göttingen, Germany.; NanoTag Biotechnologies GmbH, Göttingen, Germany., Rizzoli SO; Institute for Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany. srizzol@gwdg.de.; Center for Biostructural Imaging of Neurodegeneration, University Medical Center Göttingen, Göttingen, Germany. srizzol@gwdg.de.; Cluster of Excellence 'Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells' (MBExC), University of Göttingen, Göttingen, Germany. srizzol@gwdg.de.
Jazyk:	angličtina
Zdroj:	Nature biotechnology [Nat Biotechnol] 2024 Oct 09. Date of Electronic Publication: 2024 Oct 09.
DOI:	10.1038/s41587-024-02431-9
Abstrakt:	The attainable resolution of fluorescence microscopy has reached the subnanometer range, but this technique still fails to image the morphology of single proteins or small molecular complexes. Here, we expand the specimens at least tenfold, label them with conventional fluorophores and image them with conventional light microscopes, acquiring videos in which we analyze fluorescence fluctuations. One-step nanoscale expansion (ONE) microscopy enables the visualization of the shapes of individual membrane and soluble proteins, achieving around 1-nm resolution. We show that conformational changes are readily observable, such as those undergone by the ~17-kDa protein calmodulin upon Ca 2+ binding. ONE is also applied to clinical samples, analyzing the morphology of protein aggregates in cerebrospinal fluid from persons with Parkinson disease, potentially aiding disease diagnosis. This technology bridges the gap between high-resolution structural biology techniques and light microscopy, providing new avenues for discoveries in biology and medicine. (© 2024. The Author(s).)
Databáze:	MEDLINE
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