scholarly journals Impact of Hydrostatic Pressure on an Intrinsically Disordered Protein: A High-Pressure NMR Study of α-Synuclein

ChemBioChem ◽  
2013 ◽  
Vol 14 (14) ◽  
pp. 1754-1761 ◽  
Author(s):  
Julien Roche ◽  
Jinfa Ying ◽  
Alexander S. Maltsev ◽  
Ad Bax
Keyword(s):  
High Pressure ◽  
Hydrostatic Pressure ◽  
Protein A ◽  
Intrinsically Disordered Protein ◽  
Intrinsically Disordered ◽  
Disordered Protein ◽  
Nmr Study

scholarly journals In-Cell NMR Study of Tau and MARK2 Phosphorylated Tau

2018 ◽  
Vol 20 (1) ◽  
pp. 90 ◽  
Author(s):  
Shengnan Zhang ◽  
Chuchu Wang ◽  
Jinxia Lu ◽  
Xiaojuan Ma ◽  
Zhenying Liu ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Protein Structures ◽  
Intrinsically Disordered Protein ◽  
Phosphorylated Tau ◽  
Nmr Spectrum ◽  
Cellular Protection ◽  
Protection Mechanism ◽  
Intrinsically Disordered ◽  
Disordered Protein ◽  
Nmr Study

The intrinsically disordered protein, Tau, is abundant in neurons and contributes to the regulation of the microtubule (MT) and actin network, while its intracellular abnormal aggregation is closely associated with Alzheimer’s disease. Here, using in-cell Nuclear Magnetic Resonance (NMR) spectroscopy, we investigated the conformations of two different isoforms of Tau, Tau40 and k19, in mammalian cells. Combined with immunofluorescence imaging and western blot analyses, we found that the isotope-enriched Tau, which was delivered into the cultured mammalian cells by electroporation, is partially colocalized with MT and actin filaments (F-actin). We acquired the NMR spectrum of Tau in human embryonic kidney 293 (HEK-293T) cells, and compared it with the NMR spectra of Tau added with MT, F-actin, and a variety of crowding agents, respectively. We found that the NMR spectrum of Tau in complex with MT best recapitulates the in-cell NMR spectrum of Tau, suggesting that Tau predominantly binds to MT at its MT-binding repeats in HEK-293T cells. Moreover, we found that disease-associated phosphorylation of Tau was immediately eliminated once phosphorylated Tau was delivered into HEK-293T cells, implying a potential cellular protection mechanism under stressful conditions. Collectively, the results of our study reveal that Tau utilizes its MT-binding repeats to bind MT in mammalian cells and highlight the potential of using in-cell NMR to study protein structures at the residue level in mammalian cells.

scholarly journals Intrinsically Disordered Protein: A Thermodynamic Perspective

2015 ◽  
Vol 108 (2) ◽  
pp. 228a
Author(s):  
Jing Li ◽  
James O. Wrabl ◽  
Vincent J. Hilser
Keyword(s):  
Protein A ◽  
Intrinsically Disordered Protein ◽  
Intrinsically Disordered ◽  
Disordered Protein

Self-Assembling Micelles Based on an Intrinsically Disordered Protein Domain

2018 ◽  
Author(s):  
Sarah Klass ◽  
Matthew J. Smith ◽  
Tahoe Fiala ◽  
Jessica Lee ◽  
Anthony Omole ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Fusion Proteins ◽  
Organic Molecules ◽  
Intrinsically Disordered Protein ◽  
Protein Domain ◽  
Enzymatic Cleavage ◽  
Intrinsically Disordered ◽  
Disordered Protein ◽  
Self Assembling ◽  
Protein Tag

Herein, we describe a new series of fusion proteins that have been developed to self-assemble spontaneously into stable micelles that are 27 nm in diameter after enzymatic cleavage of a solubilizing protein tag. The sequences of the proteins are based on a human intrinsically disordered protein, which has been appended with a hydrophobic segment. The micelles were found to form across a broad range of pH, ionic strength, and temperature conditions, with critical micelle concentration (CMC) values below 1 µM being observed in some cases. The reported micelles were found to solubilize hydrophobic metal complexes and organic molecules, suggesting their potential suitability for catalysis and drug delivery applications.

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