scholarly journals NMR Characterization of Surface Receptor Protein Interactions in Live Cells Using Methylcellulose Hydrogels

2020 ◽  
Vol 132 (10) ◽  
pp. 3914-3918 ◽  
Author(s):  
Borja Mateos ◽  
Marco Sealey‐Cardona ◽  
Katja Balazs ◽  
Judith Konrat ◽  
Guenther Staffler ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Receptor Protein ◽  
Live Cells ◽  
Nmr Characterization ◽  
Surface Receptor

scholarly journals NMR Characterization of Surface Receptor Protein Interactions in Live Cells Using Methylcellulose Hydrogels

2020 ◽  
Vol 59 (10) ◽  
pp. 3886-3890 ◽  
Author(s):  
Borja Mateos ◽  
Marco Sealey‐Cardona ◽  
Katja Balazs ◽  
Judith Konrat ◽  
Guenther Staffler ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Receptor Protein ◽  
Live Cells ◽  
Nmr Characterization ◽  
Surface Receptor

scholarly journals NMR Characterization of the Interactions Between Glycosaminoglycans and Proteins

2021 ◽  
Vol 8 ◽  
Author(s):  
Changkai Bu ◽  
Lan Jin
Keyword(s):  
Protein Interactions ◽  
Dimensional Space ◽  
Three Dimensional ◽  
Gag Protein ◽  
Physiological Processes ◽  
Nmr Characterization ◽  
Mammalian Tissues ◽  
Three Dimensional Space

Glycosaminoglycans (GAGs) constitute a considerable fraction of the glycoconjugates found on cellular membranes and in the extracellular matrix of virtually all mammalian tissues. The essential role of GAG-protein interactions in the regulation of physiological processes has been recognized for decades. However, the underlying molecular basis of these interactions has only emerged since 1990s. The binding specificity of GAGs is encoded in their primary structures, but ultimately depends on how their functional groups are presented to a protein in the three-dimensional space. This review focuses on the application of NMR spectroscopy on the characterization of the GAG-protein interactions. Examples of interpretation of the complex mechanism and characterization of structural motifs involved in the GAG-protein interactions are given. Selected families of GAG-binding proteins investigated using NMR are also described.

scholarly journals Application of virus-like particles (VLP) to NMR characterization of viral membrane protein interactions

2016 ◽  
Vol 64 (3) ◽  
pp. 255-265 ◽  
Author(s):  
Aleksandar Antanasijevic ◽  
Carolyn Kingsley ◽  
Arnab Basu ◽  
Terry L. Bowlin ◽  
Lijun Rong ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Protein Interactions ◽  
Viral Membrane ◽  
Virus Like Particles ◽  
Nmr Characterization

scholarly journals Proteomic mapping in live Drosophila tissues using an engineered ascorbate peroxidase

2015 ◽  
Vol 112 (39) ◽  
pp. 12093-12098 ◽  
Author(s):  
Chiao-Lin Chen ◽  
Yanhui Hu ◽  
Namrata D. Udeshi ◽  
Thomas Y. Lau ◽  
Frederik Wirtz-Peitz ◽  
...  
Keyword(s):  
Ascorbate Peroxidase ◽  
Protein Interactions ◽  
Protein Complexes ◽  
Cell Types ◽  
Live Cells ◽  
Specific Cell ◽  
Physiological Conditions ◽  
Subcellular Domains ◽  
Endogenous Proteins

Characterization of the proteome of organelles and subcellular domains is essential for understanding cellular organization and identifying protein complexes as well as networks of protein interactions. We established a proteomic mapping platform in live Drosophila tissues using an engineered ascorbate peroxidase (APEX). Upon activation, the APEX enzyme catalyzes the biotinylation of neighboring endogenous proteins that can then be isolated and identified by mass spectrometry. We demonstrate that APEX labeling functions effectively in multiple fly tissues for different subcellular compartments and maps the mitochondrial matrix proteome of Drosophila muscle to demonstrate the power of APEX for characterizing subcellular proteomes in live cells. Further, we generate “MitoMax,” a database that provides an inventory of Drosophila mitochondrial proteins with subcompartmental annotation. Altogether, APEX labeling in live Drosophila tissues provides an opportunity to characterize the organelle proteome of specific cell types in different physiological conditions.

scholarly journals The Fluorescent Two-Hybrid Assay to Screen for Protein–Protein Interaction Inhibitors in Live Cells

2014 ◽  
Vol 19 (4) ◽  
pp. 516-525 ◽  
Author(s):  
Larisa Yurlova ◽  
Maarten Derks ◽  
Andrea Buchfellner ◽  
Ian Hickson ◽  
Marc Janssen ◽  
...  
Keyword(s):  
Small Molecules ◽  
Protein Interactions ◽  
Mammalian Cells ◽  
Live Cells ◽  
Protein Protein Interactions ◽  
Stapled Peptides ◽  
Protein Protein Interaction ◽  
Real Time Visualization ◽  
Two Hybrid

Protein–protein interactions (PPIs) are attractive but challenging targets for drug discovery. To overcome numerous limitations of the currently available cell-based PPI assays, we have recently established a fully reversible microscopy-assisted fluorescent two-hybrid (F2H) assay. The F2H assay offers a fast and straightforward readout: an interaction-dependent co-localization of two distinguishable fluorescent signals at a defined spot in the nucleus of mammalian cells. We developed two reversible F2H assays for the interactions between the tumor suppressor p53 and its negative regulators, Mdm2 and Mdm4. We then performed a pilot F2H screen with a subset of compounds, including small molecules (such as Nutlin-3) and stapled peptides. We identified five cell-penetrating compounds as potent p53–Mdm2 inhibitors. However, none exhibited intracellular activity on p53–Mdm4. Live cell data generated by the F2H assays enable the characterization of stapled peptides based on their ability to penetrate cells and disrupt p53–Mdm2 interaction as well as p53–Mdm4 interaction. Here, we show that the F2H assays enable side-by-side analysis of substances’ dual Mdm2–Mdm4 activity. In addition, they are suitable for testing various types of compounds (e.g., small molecules and peptidic inhibitors) and concurrently provide initial data on cellular toxicity. Furthermore, F2H assays readily allow real-time visualization of PPI dynamics in living cells.

Characterization of a Motile Cellular Domain Arising During the Amoebo-Flagellate Transformation of Physarum Polycephalum

1980 ◽  
Vol 38 ◽  
pp. 552-553
Author(s):  
K.I. Pagh ◽  
M.R. Adelman
Keyword(s):  
Cell Shape ◽  
Physarum Polycephalum ◽  
Slime Mold ◽  
Live Cells ◽  
Cellular Domain

Unicellular amoebae of the slime mold Physarum polycephalum undergo marked changes in cell shape and motility during their conversion into flagellate swimming cells (l). To understand the processes underlying motile activities expressed during the amoebo-flagellate transformation, we have undertaken detailed investigations of the organization, formation and functions of subcellular structures or domains of the cell which are hypothesized to play a role in movement. One focus of our studies is on a structure, termed the “ridge” which appears as a flattened extension of the periphery along the length of transforming cells (Fig. 1). Observations of live cells using Nomarski optics reveal two types of movement in this region:propagation of undulations along the length of the ridge and formation and retraction of filopodial projections from its edge. The differing activities appear to be associated with two characteristic morphologies, illustrated in Fig. 1.

NMR characterization of complex natural products: Assigning novel, proton-deficient alkaloid scaffolds

Planta Medica ◽  
2016 ◽  
Vol 81 (S 01) ◽  
pp. S1-S381
Author(s):  
KR Gustafson ◽  
STS Chan ◽  
D Milanowski
Keyword(s):  
Natural Products ◽  
Nmr Characterization
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