In-cell NMR spectroscopy is an effective tool for observing proteins at atomic resolution in their native cellular environment. However, its utility is limited by its low sensitivity and the extensive line broadening caused by nonspecific interactions in the cells, which is even more pronounced in human cells due to the difficulty of overexpressing or delivering high concentrations of isotopically labeled proteins. Here, we present a high-sensitivity tag (wPSP-6F) containing two trifluoromethyl...
[NMR paper] Toward protein NMR at physiological concentrations by hyperpolarized water-Finding and mapping uncharted conformational spaces
Toward protein NMR at physiological concentrations by hyperpolarized water-Finding and mapping uncharted conformational spaces
Nuclear magnetic resonance (NMR) spectroscopy is a key method for determining the structural dynamics of proteins in their native solution state. However, the low sensitivity of NMR typically necessitates nonphysiologically high sample concentrations, which often limit the relevance of the recorded data. We show how to use hyperpolarized water by dissolution dynamic nuclear polarization (DDNP) to acquire protein spectra at concentrations of 1 ?M within seconds and...
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08-06-2022 07:25 AM
[NMR paper] Visualizing Proteins in Mammalian Cells by 19F NMR spectroscopy
Visualizing Proteins in Mammalian Cells by 19F NMR spectroscopy
In-cell NMR spectroscopy is a powerful tool to investigate protein behavior in physiologically relevant environments. Although proven valuable for disordered proteins, we show that in commonly used 1 H- 15 N HSQC spectra of globular proteins, interactions with cellular components often broaden resonances beyond detection. This contrasts 19 F spectra in mammalian cells, in which signals are readily observed. Using several proteins, we demonstrate that surface charges and interaction with cellular...
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03-14-2022 04:46 AM
[NMR paper] Positively-Charged Tags Impede Protein Mobility in Cells as Quantified by 19F NMR.
Positively-Charged Tags Impede Protein Mobility in Cells as Quantified by 19F NMR.
Related Articles Positively-Charged Tags Impede Protein Mobility in Cells as Quantified by 19F NMR.
J Phys Chem B. 2019 May 01;:
Authors: Ye Y, Wu Q, Zheng W, Jiang B, Pielak GJ, Liu M, Li C
Abstract
Proteins are often tagged for visualization or delivery in the "sea" of other macromolecules in cells but how tags affect protein mobility remains poorly understood. Here, we employ 19F in-cell NMR to quantify the mobility of proteins with charged tags...
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05-04-2019 01:24 PM
Visualizing RNA-protein Interactions in Live Human Cells - Bioscience Technology
Visualizing RNA-protein Interactions in Live Human Cells - Bioscience Technology
http://www.bionmr.com//t3.gstatic.com/images?q=tbn:ANd9GcRoPawKAwV63VcpQ8t5e9ZjpOvT0S-FLAVv5fJ1CFHJeDztic5RkQBERPvhe9MhZmtvPn17oDdy
Bioscience Technology
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Visualizing RNA-protein Interactions in Live Human Cells
Bioscience Technology
UAlbany chemists Alan Chen Ph.D, Maksim Royzen Ph.D, and Alex Shekhtman Ph.D have combined their labs on a collaborative project aimed at visualizing RNA-protein interactions inside live human cells by using multi-dimensional NMR (nuclear ...
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06-03-2017 11:49 AM
[NMR paper] A comparison of chemical shift sensitivity of trifluoromethyl tags: optimizing resolution in (19)F NMR studies of proteins.
A comparison of chemical shift sensitivity of trifluoromethyl tags: optimizing resolution in (19)F NMR studies of proteins.
A comparison of chemical shift sensitivity of trifluoromethyl tags: optimizing resolution in (19)F NMR studies of proteins.
J Biomol NMR. 2015 Mar 27;
Authors: Ye L, Larda ST, Frank Li YF, Manglik A, Prosser RS
Abstract
The elucidation of distinct protein conformers or states by fluorine ((19)F) NMR requires fluorinated moieties whose chemical shifts are most sensitive to subtle changes in the local...
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03-31-2015 07:17 PM
A comparison of chemical shift sensitivity of trifluoromethyl tags: optimizing resolution in 19 F NMR studies of proteins
A comparison of chemical shift sensitivity of trifluoromethyl tags: optimizing resolution in 19 F NMR studies of proteins
Abstract
The elucidation of distinct protein conformers or states by fluorine (19F) NMR requires fluorinated moieties whose chemical shifts are most sensitive to subtle changes in the local dielectric and magnetic shielding environment. In this study we evaluate the effective chemical shift dispersion of a number of thiol-reactive trifluoromethyl probes N-(4-(trifluoromethyl)phenyl)acetamide (BTFMA),...
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03-27-2015 07:43 AM
[NMR paper] Development of an antibody-based, modular biosensor for 129Xe NMR molecular imaging of cells at nanomolar concentrations.
Development of an antibody-based, modular biosensor for 129Xe NMR molecular imaging of cells at nanomolar concentrations.
Development of an antibody-based, modular biosensor for 129Xe NMR molecular imaging of cells at nanomolar concentrations.
Proc Natl Acad Sci U S A. 2014 Jul 28;
Authors: Rose HM, Witte C, Rossella F, Klippel S, Freund C, Schröder L
Abstract
Magnetic resonance imaging (MRI) is seriously limited when aiming for visualization of targeted contrast agents. Images are reconstructed from the weak diamagnetic...
Visualizing Proteins in Human Cells at Near-Physiological Concentrations with*Sensitive 19F NMR Chemical Tags
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