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| Open AccessSpatial snapshots of amyloid precursor protein intramembrane processing via early endosome proteomics
Methods to assess organellar content are important. Here, Park et al develop a method for rapid isolation of early/sorting endosomes and demonstrate the application of the approach for analysis of endosomal proteomes and lipidomes, and for analysis of APP processing to Aβ via β and γ-Secretases.
- Hankum Park
- , Frances V. Hundley
- & J. Wade Harper
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Article
| Open AccessScalable multiplex co-fractionation/mass spectrometry platform for accelerated protein interactome discovery
Co-fractionation/mass spectrometry (CF/MS) allows mapping protein interactomes but efficiency and quantitative accuracy are limited. Here, the authors develop a reproducible multiplexed CF/MS method and apply it to characterize interactome rewiring in breast cancer cells.
- Pierre C. Havugimana
- , Raghuveera Kumar Goel
- & Andrew Emili
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Article
| Open AccessSpySwitch enables pH- or heat-responsive capture and release for plug-and-display nanoassembly
The SpyCatcher-SpyTag system allows protein anchoring and nanoassembly. Here, the authors engineer SpySwitch, a dually switchable Catcher which allows gentle purification of SpyTagged proteins prior to downstream applications such as the assembly of virus-like particles.
- Susan K. Vester
- , Rolle Rahikainen
- & Mark Howarth
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Article
| Open AccessA mass spectrometric method for in-depth profiling of phosphoinositide regioisomers and their disease-associated regulation
Different phosphoinositide isomers are involved in a variety of physiological and pathological processes. Here, the authors combine chiral column chromatography and mass spectrometry to measure phosphoinositide regioisomers, revealing their dynamic changes in intra- and extracellular cancer cell milieus.
- Shin Morioka
- , Hiroki Nakanishi
- & Takehiko Sasaki
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Article
| Open AccessDynamic tracking and identification of tissue-specific secretory proteins in the circulation of live mice
The in vivo identification of proteins secreted from a specific cell type or tissue remains challenging. Here, the authors develop a proximity labeling-based method to selectively label secreted proteins and combine it with proteomics to identify liver secretory proteins in mouse plasma.
- Kwang-eun Kim
- , Isaac Park
- & Jae Myoung Suh
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Article
| Open AccessUbiB proteins regulate cellular CoQ distribution in Saccharomyces cerevisiae
Coenzyme Q (CoQ) is a lipid made in the inner mitochondrial membrane with antioxidant roles throughout the cell, but regulation of its cellular distribution is unclear. Here the authors identify two proteins that have reciprocal CoQ trafficking functions to help coordinate CoQ localization in yeast.
- Zachary A. Kemmerer
- , Kyle P. Robinson
- & David J. Pagliarini
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Article
| Open Access3D projection electrophoresis for single-cell immunoblotting
Single-cell immunoblotting previously separated proteins on a polyacrylamide slab in the xy direction and was limited by throughput and sample consumption. Here the authors adapt the system to separate proteins in the z direction, allowing for closer spacing of sample wells and improved sample consumption.
- Samantha M. Grist
- , Andoni P. Mourdoukoutas
- & Amy E. Herr
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Article
| Open AccessEngineering Af1521 improves ADP-ribose binding and identification of ADP-ribosylated proteins
ADP-ribose binding macro domains facilitate the enrichment and detection of cellular ADP-ribosylation. Here, the authors generate an engineered macro domain with increased ADP-ribose affinity, improving the identification of ADP-ribosylated proteins by proteomics, western blot and immunofluorescence.
- Kathrin Nowak
- , Florian Rosenthal
- & Michael O. Hottiger
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Article
| Open AccessRaman image-activated cell sorting
Most current cell sorting methods are based on fluorescence detection with no imaging capability. Here the authors generate and use Raman image-activated cell sorting with a throughput of around 100 events per second, providing molecular images with no need for labeling.
- Nao Nitta
- , Takanori Iino
- & Keisuke Goda
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Article
| Open AccessClassification of mouse B cell types using surfaceome proteotype maps
Analysis of the cell surface proteome (surfaceome) is essential for cell classification but is technically challenging. Here the authors miniaturize and automate the Cell Surface Capture method to increase sensitivity, reproducibility and throughput, and use it to create population-specific surfaceome maps of developing mouse B cells.
- Marc van Oostrum
- , Maik Müller
- & Bernd Wollscheid
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Article
| Open AccessThe ALFA-tag is a highly versatile tool for nanobody-based bioscience applications
Epitope tags are widely used in various applications, but often lack versatility. Here, the authors introduce a small, alpha helical tag, which is recognized by a high affinity nanobody and can be used in a range of different applications, from protein purification to super-resolution imaging and in vivo detection of proteins.
- Hansjörg Götzke
- , Markus Kilisch
- & Steffen Frey
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Article
| Open AccessBrain leptin reduces liver lipids by increasing hepatic triglyceride secretion and lowering lipogenesis
Obesity is associated with leptin resistance and rising blood leptin levels while central leptin exposure may be limited. Here, the authors show that brain leptin infusion reduces hepatic lipid content in rats by increasing hepatic VLDL secretion and lowering liver de novo lipogenesis via a vagal mechanism.
- Martina Theresa Hackl
- , Clemens Fürnsinn
- & Thomas Scherer
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Article
| Open AccessSimultaneous and stoichiometric purification of hundreds of oligonucleotides
Chemically synthesized DNA oligonucleotides (oligos) require purification to remove truncated species. Here, the authors developed a high-throughput method for oligo purification that also normalises the concentrations of the oligos in the final samples.
- Alessandro Pinto
- , Sherry X. Chen
- & David Yu Zhang
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Article
| Open AccessA photoelectrochemical platform for the capture and release of rare single cells
Many cell capture systems exist but the characterisation and controlled release of single cells is a challenge. Here, the authors report on the development of a duel trigger release system using a combination of photo and electro triggers to allow for light based analysis without unwanted release.
- Stephen G. Parker
- , Ying Yang
- & J. Justin Gooding
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Article
| Open AccessCombining laser capture microdissection and proteomics reveals an active translation machinery controlling invadosome formation
Invadosomes degrade extracellular matrix and facilitate cell invasion but their molecular composition is not fully understood. Here, the authors combine laser capture and mass spectrometry to map the proteome of invadosomes, showing that they rely on internal translational activity to maintain their structure.
- Zakaria Ezzoukhry
- , Elodie Henriet
- & Frédéric Saltel
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Article
| Open AccessAn enrichment method based on synergistic and reversible covalent interactions for large-scale analysis of glycoproteins
Understanding the functions of protein glycosylation critically depends on methods to efficiently enrich glycoproteins from complex samples. Here, the authors develop a strategy using dendrimer-conjugated benzoboroxole to enhance glycopeptide enrichment, providing the basis for more comprehensive glycoprotein analyses.
- Haopeng Xiao
- , Weixuan Chen
- & Ronghu Wu
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Article
| Open AccessHydrogen bond based smart polymer for highly selective and tunable capture of multiply phosphorylated peptides
Capture of low-abundance multiply phosphorylated peptides (MPPs) is difficult due to limitation of enrichment materials and their interactions with phosphates. Here the authors show, a smart polymer driven by specific but tunable hydrogen bonding interactions can differentially complex with MPPs, singly phosphorylated and non-modified peptides.
- Guangyan Qing
- , Qi Lu
- & Taolei Sun
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Article
| Open AccessEfficient protein production inspired by how spiders make silk
The properties of many transmembrane or aggregation-prone proteins make them difficult to recombinantly express. Here the authors use a modified N-terminal domain of a spider silk protein to express and purify several difficult to express proteins at levels considerably higher than with conventional tags.
- Nina Kronqvist
- , Médoune Sarr
- & Jan Johansson
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Article
| Open AccessTrapping mammalian protein complexes in viral particles
A large portion of the proteome carries out its cellular function as part of macromolecular complexes. Here the authors describe Virotrap, a novel lysis-free approach for the isolation and identification of biologically relevant protein-protein and small molecule-protein interactions.
- Sven Eyckerman
- , Kevin Titeca
- & Jan Tavernier
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Article
| Open AccessSerial interactome capture of the human cell nucleus
RNA-binding proteins are involved in the posttranscriptional regulation of a large number of cellular processes and several recent studies have sought to describe the extent of the RNA-binding proteome. Here, Conrad et al. describe serIC, a stringent approach they apply towards defining the RNA-binding proteome of the mammalian nucleus.
- Thomas Conrad
- , Anne-Susann Albrecht
- & Ulf Andersson Ørom
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Article |
A quantitative telomeric chromatin isolation protocol identifies different telomeric states
The protein composition of telomeres changes during development, aging, tumourigenesis and in telomere syndromes. Here, the authors develop a quantitative telomeric chromatin isolation protocol (QTIP) to analyse and quantitatively compare telomeric chromatin of different cell populations.
- Larissa Grolimund
- , Eric Aeby
- & Joachim Lingner
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Rotational separation of non-spherical bioparticles using I-shaped pillar arrays in a microfluidic device
Microfluidic separation devices are usually designed for spherical particles, but many biological particles are non-spherical; for example, red blood cells and bacteria. Using I-shaped pillar designs, Zhang et al. demonstrate a better sorting capability for non-spherical particles.
- Kerwin Kwek Zeming
- , Shashi Ranjan
- & Yong Zhang
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| Open AccessLarge-scale single-chirality separation of single-wall carbon nanotubes by simple gel chromatography
Large-scale separation of single-wall carbon nanotubes into populations of single chirality is a significant challenge in the practical application of nanotubes. Now, using multicolumn gel chromatography, the large-scale separation of 13 different carbon nanotube species is achieved.
- Huaping Liu
- , Daisuke Nishide
- & Hiromichi Kataura