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  • Journal article

  • A database of accurate electrophoretic migration patterns for human proteins in cell lines

    • Authors : Mylonas, Roman (2023)

  • Native molecular weight (MW) is one of the defining features of proteins. Denaturing gel electrophoresis (SDS-PAGE) is a very popular technique for separating proteins and determining their MW. Coupled with antibody-based detection, SDS-PAGE is widely applied for protein identification and quantitation. Yet, electrophoresis is poorly reproducible and the MWs obtained are often inaccurate. This hampers antibody validation and negatively impacts the reliability of western...
  • Journal Article

  • Evolving spike-protein N-glycosylation in SARS-CoV-2 variants

    • Authors : Baboo, Sabyasachi (2023)

  • It has been three years since SARS-CoV-2 emerged and the world plunged into a “once in a century” pandemic. Since then, multiple waves of infection have swept through the human population, led by variants that were able to evade any acquired immunity. The co-evolution of SARS-CoV-2 variants with human immunity provides an excellent opportunity to study the interaction between viral pathogens and their human hosts. The heavily N-gl...
  • Journal article

  • The role of glutathione in periplasmic redox homeostasis and oxidative protein folding in Escherichia coli

    • Authors : Knoke, Lisa R. (2023)

  • The thiol redox balance in the periplasm of E. coli depends on the DsbA/B pair for oxidative 14 power and the DsbC/D system as its complement for isomerization of non-native disulfides. 15 While the standard redox potentials of those systems are known, the in vivo redox potential 16 imposed onto protein thiol disulfide pairs in the periplasm remains unknown. Here, we used 17 genetically encoded redox probes (roGFP2 ...
  • Journal article

  • Torsion angles to map and visualize the conformational space of a protein

    • Authors : Ginn, Helen Mary (2023)

  • Present understanding of protein structure dynamics trails behind that of static structures. A torsionangle based approach, called representation of protein entities (RoPE), derives an interpretable conformational space which correlates with data collection temperature, resolution and reaction coordinate. For more complex systems, atomic coordinates fail to separate functional conformational states, which are still preserved by torsion angle-derived space. This indicates...
  • Journal article

  • Ambient temperature crystal structure of Escherichia coli CyaY protein displays alternate conformation

    • Authors : Shafiei, Alaleh (2023)

  • Frataxin is a 23 KDa mitochondrial iron-binding protein that is involved in biogenesis of iron sulfur clusters. A deficiency in frataxin leads to Friedreich's ataxia, a progressive neurodegenerative disorder. The bacterial ortholog of eukaryotic mitochondrial frataxin, CyaY, is thought to play a role in iron sulfur cluster assembly as an iron supplier, making it an important target for study. Here, we present the first ambient tem...
  • Journal article

  • Chemoproteomics-Enabled Discovery of a Covalent Molecular Glue Degrader Targeting NF-κB

    • Authors : King, Elizabeth A. (2023)

  • Targeted protein degradation using heterobifunctional Proteolysis-Targeting Chimeras (PROTACs) or molecular glues has arisen as a powerful therapeutic modality for degrading disease targets. While PROTAC design is becoming more modular and straightforward, the discovery of novel molecular glue degraders has been more challenging. While several recent studies have showcased phenotypic screening and counter-screening approaches to discover new molecular glue degraders, me...
  • Journal article

  • Lipid nanodiscs as a template for high-resolution cryo-EM structures of peripheral membrane proteins

    • Authors : Cannon, Kevin S. (2023)

  • processes such as signal transduction, membrane trafficking, and autophagy. Transient binding to the membrane has a profound impact on protein function, serving to induce conformational changes and alter biochemical and biophysical parameters by increasing the local concentration of factors and restricting diffusion to two dimensions. Despite the centrality of the membrane in serving as a template for cell biology, there are few reported highresolution stru...
  • 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&...