Isolation, separation and purification

  • Article
    | Open Access

    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
  • Article
    | Open Access

    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
  • Article
    | Open Access

    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
  • Article
    | Open Access

    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
  • Article
    | Open Access

    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
  • Article
    | Open Access

    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
  • Article
    | Open Access

    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
  • Article
    | Open Access

    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
  • Article
    | Open Access

    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
  • Article
    | Open Access

    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
  • Article
    | Open Access

    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
  • Article
    | Open Access

    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
  • Article
    | Open Access

    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
  • Article
    | Open Access

    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
  • Article
    | Open Access

    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