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Article
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Filamin A organizes γ‑aminobutyric acid type B receptors at the plasma membrane
GABAB receptors mediate the effects of the main inhibitory neurotransmitter in the brain. Here, authors identify the cytoskeletal protein filamin A as a key player that controls the exact location and function of GABAB receptors at the cell surface.
- Marie-Lise Jobin
- , Sana Siddig
- & Davide Calebiro
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| Open Access
Improved immunoassay sensitivity and specificity using single-molecule colocalization
A major challenge of enzyme-linked immunosorbent assays is discriminating true signal from non-specific binding. Here the authors present a Single-Molecule Colocalization Assay (SiMCA) which eliminates such effects, enabling reproducible detection of picomolar protein concentrations.
- Amani A. Hariri
- , Sharon S. Newman
- & H. Tom Soh
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| Open Access
Structural basis of rapid actin dynamics in the evolutionarily divergent Leishmania parasite
The authors report here the structure-function analysis of highly divergent actin from Leishmania parasite. The study reveals remarkably rapid dynamics of parasite actin as well as the underlying molecular basis, thus providing insight into evolution of the actin cytoskeleton.
- Tommi Kotila
- , Hugo Wioland
- & Pekka Lappalainen
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Article
| Open Access
Two-colour single-molecule photoinduced electron transfer fluorescence imaging microscopy of chaperone dynamics
Revealing mechanisms of complex protein machines requires simultaneous exploration of multiple structural coordinates. Here the authors report two-colour fluorescence microscopy combined with photoinduced electron transfer probes to simultaneously detect two structural coordinates in single protein molecules.
- Jonathan Schubert
- , Andrea Schulze
- & Hannes Neuweiler
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Article
| Open Access
Planar photonic chips with tailored angular transmission for high-contrast-imaging devices
The authors design a planar photonic chip with several multilayers of photonic band gaps and a region of dielectric nanoparticles for tailored angular transmission. They use it as sample substrate for high-contrast darkfield and total internal reflection microscopy on a conventional microscope.
- Yan Kuai
- , Junxue Chen
- & Douguo Zhang
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Article
| Open Access
Simultaneous orientation and 3D localization microscopy with a Vortex point spread function
Molecular orientation is often ignored during single-molecule localisation microscopy. Here, the authors use a Vortex point spread function in order to simultaneously estimate the 3D position, dipole orientation and degree of rotational constraint, within 30% of the Cramér-Rao bound limit.
- Christiaan N. Hulleman
- , Rasmus Ø. Thorsen
- & Bernd Rieger
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Article
| Open Access
Mitochondrial adaptor TRAK2 activates and functionally links opposing kinesin and dynein motors
Mitochondrial transport toward both the plus- and minus-ends of microtubules is mediated by motor proteins linked to mitochondria by TRAK adaptor proteins. Here the authors investigate the role of TRAK2 as a bidirectional motor adaptor, and propose a model where TRAK2 coordinates the activities of opposing kinesin-1 and cytoplasmic dynein motors as a single interdependent motor complex.
- Adam R. Fenton
- , Thomas A. Jongens
- & Erika L. F. Holzbaur
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| Open Access
Redefining the specificity of phosphoinositide-binding by human PH domain-containing proteins
Pleckstrin homology (PH) domains are presumed to bind phosphoinositides (PIPs) but only few specific interactions are known. Using TIRF microscopy based assays, functional studies and an experimentally validated prediction algorithm, the authors show that specific PIP binding is widespread among human PH domains.
- Nilmani Singh
- , Adriana Reyes-Ordoñez
- & Jie Chen
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Article
| Open Access
Fast widefield scan provides tunable and uniform illumination optimizing super-resolution microscopy on large fields
Uniform illumination is a prerequisite for quantitative analyses in both classical fluorescence microscopy and single molecule localisation microscopy. Here, the authors introduce ASTER, an illumination technique that generates uniform illumination over large and adaptable fields of view, compatible with epifluorescence, HiLo and TIRF illumination schemes.
- Adrien Mau
- , Karoline Friedl
- & Sandrine Lévêque-Fort
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Article
| Open Access
Direct supercritical angle localization microscopy for nanometer 3D superresolution
Supercritical angle localisation microscopy (SALM) allows the z-positions of single fluorophores to be extracted from the intensity of supercritical angle fluorescence. Here the authors improve the z-resolution of SALM, and report nanometre isotropic localisation precision on DNA origami structures.
- Anindita Dasgupta
- , Joran Deschamps
- & Jonas Ries
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Article
| Open Access
Automatic classification and segmentation of single-molecule fluorescence time traces with deep learning
Traces from single-molecule fluorescence microscopy (SMFM) experiments exhibit photophysical artifacts that typically make analysis time-consuming. Here, the authors have developed an easily accessible software, AutoSiM, for two distinct applications of deep learning to the efficient processing of SMFM time traces.
- Jieming Li
- , Leyou Zhang
- & Nils G. Walter
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Article
| Open Access
Mitochondria-adaptor TRAK1 promotes kinesin-1 driven transport in crowded environments
Intracellular trafficking of organelles is driven by kinesin-1 stepping along microtubules, but crowding conditions impede kinesin-1 motility. Here authors demonstrate that TRAK1, an adaptor protein essential for mitochondrial trafficking, activates kinesin-1 and increases robustness of kinesin-1 stepping on crowded microtubule surfaces.
- Verena Henrichs
- , Lenka Grycova
- & Zdenek Lansky
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Article
| Open Access
Direct observation of dynamic protein interactions involving human microtubules using solid-state NMR spectroscopy
Microtubule (MT) organization is regulated by many microtubule-associated proteins (MAPs) that contain intrinsically disordered regions. Here authors produce [13C, 15N] labeled, functional microtubules from human cells for solid-state NMR which allows studying MAP-MT interactions.
- Yanzhang Luo
- , ShengQi Xiang
- & Marc Baldus
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Article
| Open Access
Fermi level-tuned optics of graphene for attocoulomb-scale quantification of electron transfer at single gold nanoparticles
Measurement of single-molecule level electron transfer is restricted by detection limits in nanoampere to picoampere ranges. Here the authors develop graphene-based electrochemical microscopy to attain an attoampere-level detection limit for faraday current at single nanoparticles.
- Qing Xia
- , Zixuan Chen
- & Jun-Jie Zhu
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Article
| Open Access
The companion of cellulose synthase 1 confers salt tolerance through a Tau-like mechanism in plants
The Arabidopsis CC1 protein maintains microtubule array stability and cellulose synthesis during salt stress. Here the authors show that CC1 engages microtubules via an intrinsically disordered N-terminus that suggests it controls microtubule dynamics in a similar way to the mammalian Tau protein.
- Christopher Kesten
- , Arndt Wallmann
- & Staffan Persson
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Article
| Open Access
Multi-color live-cell super-resolution volume imaging with multi-angle interference microscopy
3D super-resolution imaging of dynamic processes in live cells is still challenging, especially in a large field of view. Here the authors combine SIM with multi-angle evanescent light illumination and achieve improved lateral and axial resolution, with stack acquisition time in the range of 1–2 s.
- Youhua Chen
- , Wenjie Liu
- & Xu Liu
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Article
| Open Access
Mechanochemical feedback control of dynamin independent endocytosis modulates membrane tension in adherent cells
Plasma membrane tension is an important factor that regulates many key cellular processes. Here authors show that a specific dynamin-independent endocytic pathway is modulated by changes in tension via the mechano-transducer vinculin.
- Joseph Jose Thottacherry
- , Anita Joanna Kosmalska
- & Satyajit Mayor
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Article
| Open Access
Processive chitinase is Brownian monorail operated by fast catalysis after peeling rail from crystalline chitin
Processive chitinase is a linear molecular motor which moves on the surface of crystalline chitin. Here authors use single-molecule imaging, X-ray crystallography and simulations on chitinase A (SmChiA) and show that Brownian motion along the single chitin chain is rectified forward by substrate-assisted catalysis.
- Akihiko Nakamura
- , Kei-ichi Okazaki
- & Ryota Iino
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| Open Access
In vivo single-molecule imaging of syntaxin1A reveals polyphosphoinositide- and activity-dependent trapping in presynaptic nanoclusters
Syntaxin1A (Sx1A) is organized in nanoclusters in neurosecretory cells but how these nanoclusters are affected by neurotransmitter release in a living organism is unknown. Here the authors perform single molecule imaging analysis in live fly larvae and show that the lateral diffusion and trapping of Sx1A in nanoclusters are altered by synaptic activity.
- Adekunle T. Bademosi
- , Elsa Lauwers
- & Frédéric A. Meunier
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Article
| Open Access
Real-time fluorescence imaging with 20 nm axial resolution
Mapping the nanoscale height and dynamics of structures within the cell is difficult. Here the authors present a two-wavelength total internal reflection fluorescence method to perform real-time imaging with nanometre axial resolution using a conventional microscope.
- Daniel R. Stabley
- , Thomas Oh
- & Khalid Salaita
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Article
| Open Access
Single-molecule imaging of a three-component ordered actin disassembly mechanism
The roles of Coronin, Cofilin and AIP1 in promoting actin disassembly have not been well understood. Here using single-molecule fluorescence imaging, Jansen et al. show that the three proteins act together in a coordinated, temporal pathway to induce rapid severing and disassembly of actin filaments.
- Silvia Jansen
- , Agnieszka Collins
- & Bruce L. Goode
Dual-color DNA-PAINT single-particle tracking enables extended studies of membrane protein interactions