We have performed dynamic nuclear polarization (DNP) experiments at liquid helium temperatures using a low-power (<70 mW) solid-state diode microwave source at 200 GHz-the electron paramagnetic resonance frequency of stable radicals at 7 T. We employed a home-built Alderman-Grant probe for the detection of ¹H NMR signal at 300 MHz, as such coils are well suited for higher frequency NMR detection. The Alderman-Grant coil is inductively coupled to the rest of the radiofrequency (rf) circuit, whose design allows probe components to be placed away from the sample area, and also enables easy switching of coils with different diameters and resonance frequencies. We have tested our DNP instrument on a frozen nitroxide model system consisting of 4-Amino TEMPO dissolved in a glycerol:water mixture. The largest nuclear spin polarization observed was 61 ± 2% with a sample containing 20 mM 4-Amino TEMPO dissolved in deuterated glycerol (d-glycerol):D₂O:H₂O (50:40:10), amounting to record polarization measured to date at an easily amenable temperature of 4 K.
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http://dx.doi.org/10.1016/j.jmr.2012.05.017 | DOI Listing |
Protein Sci
February 2025
Department of Chemistry and Biochemistry, Center for RNA Biology, The Ohio State University, Columbus, Ohio, USA.
Loz1 is a zinc-responsive transcription factor in fission yeast that maintains cellular zinc homeostasis by repressing the expression of genes required for zinc uptake in high zinc conditions. Previous deletion analysis of Loz1 found a region containing two tandem CH zinc-fingers and an upstream "accessory domain" rich in histidine, lysine, and arginine residues to be sufficient for zinc-dependent DNA binding and gene repression. Here we report unexpected biophysical properties of this pair of seemingly classical CH zinc fingers.
View Article and Find Full Text PDFBiomark Res
January 2025
BK21 FOUR KNU Convergence Educational Program of Biomedical Sciences for Creative Future Talents, Department of Biomedical Sciences, School of Medicine, Kyungpook National University, Daegu, 41944, Korea.
Macrophages are pivotal in the body's defense and response to inflammation. They are present in significant numbers and are widely implicated in various diseases, including cancer. While molecular and histological techniques have advanced our understanding of macrophage biology, their precise function within the cancerous microenvironments remains underexplored.
View Article and Find Full Text PDFBiochemistry (Mosc)
December 2024
Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, 119991, Russia.
Nuclear export protein (NEP) of the influenza A virus, being one of the key components of the virus life cycle, is a promising model for studying characteristics of formation of amyloids by viral proteins. Using atomic force microscopy, comparative study of aggregation properties of the recombinant NEP variants, including the protein of natural structure, as well as modified variants with N- and C-terminal affinity His-tags, was carried out. All protein variants under physiological conditions are capable of forming aggregates of various morphologies: micelle-like nanoparticles, flexible protofibrils, rigid amyloid fibrils, etc.
View Article and Find Full Text PDFBiol Psychiatry
January 2025
Institute of Biology Paris-Seine, laboratory Neuroscience Paris-Seine, CNRS, INSERM, Sorbonne Université, UPMC Université Paris 06 F-75005, Paris, France. Electronic address:
Background: The persistence of cocaine-evoked adaptations relies on gene regulations within the reward circuit, especially in the ventral striatum (i.e., nucleus accumbens (NAc)).
View Article and Find Full Text PDFJ Biol Chem
January 2025
Department of Chemistry and Biochemistry, Ohio University, Athens, OH 45701, USA; Molecular and Cellular Biology Program, Ohio University, Athens, OH 45701, USA; Edison Biotechnology Institute, Ohio University, Athens, OH 45701, USA.
Somatic Y537S and D538G mutations within the estrogen receptor alpha ligand-binding domain (ERα-LBD) have been linked to enhanced cell proliferation, survival, and metastases in ER-positive breast cancers. Such mutations are thought to confer ligand-independent receptor activation by increasing the flexibility of helix 12 (H12), a segment within the ERα-LBD that acts as a dynamic regulator of ERα activity. We employed bipartite tetracysteine display coupled with the biarsenical profluorophore FlAsH-EDT to monitor ligand-independent structural transitions of H12 in ERα-LBDs that include Y537S or D538G mutations.
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