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  • Journal article

  • Intracellular biotransformation and disposal mechanisms of magnetosomes in macrophages and cancer cells

    • Authors :  Gandarias, L. (2023)

  • Given their potential clinical applications for the diagnosis and treatment of cancer, it is essential to understand what becomes of them once they are within the body. With this aim, here we have followed the intracellular long-term fate of magnetosomes in two cell types: cancer cells (A549 cell line), because they are the actual target for the therapeutic activity of the magnetosomes, and macrophages (RAW 264.7 cell line), ...
  • Journal article

  • Biophysical characterization and analysis of a mesophilic chorismate mutase from B. pumilus

    • Authors : Wilkins, Ryan Scott  (2023)

  • Chorismate mutases have extensively been used as computational benchmarking systems for enzyme catalysis, yet the roles entropy and enthalpy play in catalysis are still not fully understood. Thus, it is important to better understand these enzymes for potential research or industrial applications. Here, we report the first crystal structure and kinetic characterization of a chorismate mutase from Bacillus pumilus. This enzyme exhibits a high degree of similar...
  • Journal article

  • Structure of the ceramide-bound SPOTS complex

    • Authors : Schäfer, an-Hannes (2023)

  • Sphingolipids are structural membrane components that also function in cellular stress 32 responses. The serine palmitoyl-transferase (SPT) catalyzes the rate limiting step in 33 sphingolipid biogenesis. Its activity is tightly regulated through multiple binding part-34 ners, including Tsc3, Orm proteins, ceramides, and the phosphatidylinositol-4-phos-35 phate (PI4P) phosphatase Sac1. The structural organization and regulatory mecha-36 nisms of this complex are not yet understood.
  • Journal article

  • Rdh54 stabilizes Rad51 at displacement loop intermediates to regulate genetic exchange between chromosomes

    • Authors : Keymakh, Margaret (2023)

  • Homologous recombination (HR) is a double-strand break DNA repair pathway that preserves chromosome structure. To repair the damaged recipient DNA, HR requires an intact donor DNA sequence located elsewhere in the genome. After the double-strand break is repaired, DNA sequence information can be transferred between donor and recipient DNA molecules through different mechanisms, including DNA crossovers that form between homologous chromosomes. Regulating...
  • Journal Article

  • Label-free Quantification of Host-Cell Protein Impurity in a Recombinant Hemoglobin Reference Material

    • Authors : Henrion, André (2023)

  • Quantitative analysis depends on pure-substance primary calibrators with known mass fractions of impurity. Here, label-free quantification (LFQ) is being evaluated as a readily available, reliable method for determining the mass fraction of host-cell proteins (HCPs) in bioengineered proteins. For example, hemoglobin-A2 (HbA2) is being used as obtained through overexpression in E.coli. Two different materials had been produced: natural, and U-15N-labeled HbA2. For quantification&...
  • Journal article

  • Cooperativity between Cas9 and hyperactive AID establishes broad and diversifying mutational footprints in base editors

    • Authors : Berríos, Kiara N.  (2023)

  • The partnership of DNA deaminase enzymes with CRISPR-Cas nucleases is now a well-established method to enable targeted genomic base editing. However, an understanding of how Cas9 and DNA deaminases collaborate to shape base editor (BE) outcomes has been lacking. Here, we support a novel mechanistic model of base editing by deriving a range of hyperactive activation-induced deaminase (AID) base editors (hBEs) and exploiting their characteristic&#...
  • Journal article

  • Molecular basis of neurodevelopmental disorder-causing mutation in nonsense-mediated mRNA decay factor UPF3B

    • Authors : Bufton, Joshua C. (2023)

  • UPF3B is a key nonsense-mediated mRNA decay (NMD) factor required for surveillance of mRNA and regulation of eukaryotic gene expression. Mutations in UPF3B cause intellectual disability. The underlying molecular mechanisms remain unexplored as the mutations lie in an uncharacterized region of UPF3B. Here, we show that UPF3B shares structural and functional homology to the Drosophila Behavior/Human Splicing protein family comprising an RNA-recognition motif-like domain&...
  • Journal Article

  • Structural Basis for the Phase Separation of the Chromosome Passenger Complex

    • Authors : Bryan, Nikaela W. (2023)

  • Membraneless intracellular compartmentalization is central to a long and growing list of biochemical transactions at diverse sub-cellular locations 1. Proteins are by-and-large the drivers of the formation of these compartments, but there is debate about whether their underpinnings are either low-affinity/low-specificity interactions yielding phase separation 2 or multivalent site-specific interactions 3. By its very nature, ascertaining the former type of &#x...
  • Journal Article

  • tRNA m1G9 modification depends on substrate-specific RNA conformational changes induced by the methyltransferase Trm10

    • Authors : E strassler, sarah (2023)

  • The methyltransferase Trm10 modifies a subset of tRNAs on the base N1 position of the 9th nucleotide in the tRNA core. Trm10 is conserved throughout Eukarya and Archaea, and mutations in the human gene (TRMT10A) have been linked to neurological disorders such as microcephaly and intellectual disability, as well as defects in glucose metabolism. Of the 26 tRNAs in yeast with guanosine at position 9, only 14 are substrates ...