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  • Journal Article

  • Site-Specific Introduction of Alanines for the NMR Investigation of Low-Complexity Regions and Large Biomolecular Assemblies

    • Authors : Elena-Real, Carlos A.  (2023)

  • NMR studies of large biomolecular machines and highly repetitive proteins remain challenging due to the difficulty of assigning signals to individual nuclei. Here, we present an efficient strategy to address this challenge by engineering a Pyrococcus horikoshii tRNA/alanyl-tRNA synthetase pair that enables the incorporation of up to three isotopically labeled alanine residues in a site-specific manner using in vitro protein expression. We have d...
  • Journal article

  • Structural basis for binding of Smaug to the GPCR Smoothened and to the germline inducer Oskar

    • Authors : Kubíková, Jana (2023)

  • Drosophila Smaug and its orthologs comprise a family of mRNA repressor proteins that exhibit various functions during animal development. Smaug proteins contain a characteristic RNA-binding sterile-a motif (SAM) domain and a conserved but uncharacterized N-terminal domain (NTD). Here, we resolved the crystal structure of the NTD of the human SAM domain-containing protein 4A (SAMD4A, a.k.a. Smaug1) to 2.0 Å resolution, which revealed its composi...
  • Journal article

  • An alphacoronavirus polymerase structure reveals conserved co-factor functions

    • Authors : Anderson, Thomas K. (2023)

  • Coronaviruses are a diverse subfamily of viruses containing pathogens of humans and animals. This subfamily of viruses replicates their RNA genomes using a core polymerase complex composed of viral non-structural proteins: nsp7, nsp8 and nsp12. Most of our understanding of coronavirus molecular biology comes from the betacoronaviruses like SARS-CoV and SARSCoV- 2, the latter of which is the causative agent of COVID-19. In...
  • Industry article

  • Characterisation of the SUF FeS cluster machinery in the amitochondriate eukaryote Monocercomonoides exilis

    • Authors : Diaz, Priscila Peña (2023)

  • Monocercomonoides exilis is the first eukaryotic organism described as a complete amitochondriate,24 yet it shares common features with heterotrophic anaerobic/microaerophilic protists, some of which25 bear divergent mitochondrion-related organelles or MROs. It has been postulated that the retention26 of these organelles stems from their involvement in the assembly of essential cytosolic and nuclear27 FeS proteins, whose maturation requires the evolutionarily conse...
  • Journal article

  • Arachidonic acid does not grease the exocytotic SNARE machinery

    • Authors : Fasshauer, Dirk (2023)

  • It has been proposed that arachidonic acid stimulates neurite outgrowth by specifically activating syntaxin 3, a SNARE proteins that mediates the fusion of transport vesicles with the plasma membrane and is thus instrumental for the insertion of new membrane needed for cell growth and proliferation. Considering the important role of arachidonic acid and other polyunsaturated fatty acids for human health, these findings would have wide implications for ev...
  • Journal article

  • Solid Phase Synthesis of Fluorosulfate Containing Macrocycles for Chemoproteomic Workflows

    • Authors : Appleby, Franco F; Abegg, Daniel (2023)

  • Macrocyclic peptides are attractive for chemoproteomic applications due to their modular synthesis and potential for high target selectivity. We describe a solid phase synthesis method for the efficient generation of libraries of small macrocycles that contain an electrophile and alkyne handle. The modular synthesis produces libraries that can be directly screened using simple SDS-PAGE readouts and then optimal lead molecules applied to proteomic ana...
  • Journal article

  • Epigenetic mechanisms to propagate histone acetylation by p300/CBP

    • Authors : Kikuchi, Masaki  (2023)

  • Histone acetylation is important for the activation of gene transcription but little is known 14 about its direct ‘read/write’ mechanisms. Here, we report cryo-electron microscopy structures 15 in which a p300/CBP multidomain monomer recognizes histone H4 N-terminal tail (NT) 16 acetylation (ac) in a nucleosome and acetylates non-H4 histone NTs within the same 17 nucleosome. p300/CBP not only recognized H4NTac via the bromodomain pocke...