High-resolution proton and laser photochemically induced dynamic nuclear polarization NMR studies of cation binding to bovine .alpha.-lactalbumin

Biochemistry ◽  
1987 ◽  
Vol 26 (18) ◽  
pp. 5769-5774 ◽  
Author(s):  
Lawrence J. Berliner ◽  
Keiko Koga ◽  
Hiroyasu Nishikawa ◽  
Julie E. Scheffler
Keyword(s):  
High Resolution ◽  
Dynamic Nuclear Polarization ◽  
Nuclear Polarization ◽  
Cation Binding ◽  
Nmr Studies ◽  
Alpha Lactalbumin

scholarly journals In Situ Detection of Endogenous HIV Activation by Dynamic Nuclear Polarization NMR and Flow Cytometry

2020 ◽  
Vol 21 (13) ◽  
pp. 4649
Author(s):  
Sarah A. Overall ◽  
Lauren E. Price ◽  
Brice J. Albert ◽  
Chukun Gao ◽  
Nicholas Alaniva ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Dynamic Nuclear Polarization ◽  
Nuclear Polarization ◽  
Fluorescent Protein ◽  
Human Cell Line ◽  
Cellular Heterogeneity ◽  
Nmr Studies ◽  
Cellular Processes ◽  
Green Fluorescent ◽  
In Situ Detection

We demonstrate for the first time in-cell dynamic nuclear polarization (DNP) in conjunction with flow cytometry sorting to address the cellular heterogeneity of in-cell samples. Utilizing a green fluorescent protein (GFP) reporter of HIV reactivation, we correlate increased 15N resonance intensity with cytokine-driven HIV reactivation in a human cell line model of HIV latency. As few as 10% GFP+ cells could be detected by DNP nuclear magnetic resonance (NMR). The inclusion of flow cytometric sorting of GFP+ cells prior to analysis by DNP-NMR further boosted signal detection through increased cellular homogeneity with respect to GFP expression. As few as 3.6 million 15N-labeled GFP+ cells could be readily detected with DNP-NMR. Importantly, cell sorting allowed for the comparison of cytokine-treated GFP+ and GFP− cells in a batch-consistent way. This provides an avenue for normalizing NMR spectral contributions from background cellular processes following treatment with cellular modulators. We also demonstrate the remarkable stability of AMUPol (a nitroxide biradical) in Jurkat T cells and achieved in-cell enhancements of 46 with 10 mM AMUPol, providing an excellent model system for further in-cell DNP-NMR studies. This represents an important contribution to improving in-cell methods for the study of endogenously expressed proteins by DNP-NMR.

scholarly journals Probing Protein Structure by Solvent Perturbation of NMR Spectra. A Comparison with Photochemically Induced Dynamic Nuclear Polarization Techniques Applied to Native alpha-Lactalbumin

1995 ◽  
Vol 227 (1-2) ◽  
pp. 78-86 ◽  
Author(s):  
Sabina Improta ◽  
Henriette Molinari ◽  
Annalisa Pastore ◽  
Roberto Consonni ◽  
Lucia Zetta
Keyword(s):  
Protein Structure ◽  
Dynamic Nuclear Polarization ◽  
Nmr Spectra ◽  
Nuclear Polarization ◽  
Alpha Lactalbumin

scholarly journals Dynamic Nuclear Polarization of Biomembrane Assemblies

Biomolecules ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1246
Author(s):  
Nhi T. Tran ◽  
Frédéric Mentink-Vigier ◽  
Joanna R. Long
Keyword(s):  
Membrane Proteins ◽  
Dynamic Nuclear Polarization ◽  
Nuclear Polarization ◽  
Atomic Scale ◽  
Nuclear Spins ◽  
Nmr Studies ◽  
Nmr Characterization ◽  
Recent Developments ◽  
Lipid Assemblies

While atomic scale structural and dynamic information are hallmarks of nuclear magnetic resonance (NMR) methodologies, sensitivity is a fundamental limitation in NMR studies. Fully exploiting NMR capabilities to study membrane proteins is further hampered by their dilution within biological membranes. Recent developments in dynamic nuclear polarization (DNP), which can transfer the relatively high polarization of unpaired electrons to nuclear spins, show promise for overcoming the sensitivity bottleneck and enabling NMR characterization of membrane proteins under native-like conditions. Here we discuss fundamental aspects of DNP-enhanced solid-state NMR spectroscopy, experimental details relevant to the study of lipid assemblies and incorporated proteins, and sensitivity gains which can be realized in biomembrane-based samples. We also present unique insights which can be gained from DNP measurements and prospects for further development of the technique for elucidating structures and orientations of membrane proteins in native lipid environments.

scholarly journals Efficient cross-effect dynamic nuclear polarization without depolarization in high-resolution MAS NMR

2017 ◽  
Vol 8 (12) ◽  
pp. 8150-8163 ◽  
Author(s):  
Frédéric Mentink-Vigier ◽  
Guinevere Mathies ◽  
Yangping Liu ◽  
Anne-Laure Barra ◽  
Marc A. Caporini ◽  
...  
Keyword(s):  
High Resolution ◽  
Dynamic Nuclear Polarization ◽  
Nuclear Polarization ◽  
Mas Nmr ◽  
Cross Effect ◽  
Nmr Sensitivity

The mixed trityl-TEMPO biradical TEMTriPol-1 provides excellent MAS NMR sensitivity with DNP while avoiding nuclear depolarization.

Investigation of a naphthalene pitch by high-resolution solid-state dynamic nuclear polarization

1996 ◽  
Vol 6 (2) ◽  
pp. 127-133 ◽  
Author(s):  
Jianwei Zhou ◽  
Baolian Yang ◽  
Jianzhi Hu ◽  
Hongbing Hu ◽  
Liyun Li ◽  
...  
Keyword(s):  
Solid State ◽  
High Resolution ◽  
Dynamic Nuclear Polarization ◽  
Nuclear Polarization ◽  
State Dynamic

scholarly journals High-resolution Overhauser dynamic nuclear polarization enhanced proton NMR spectroscopy at low magnetic fields

2020 ◽  
Vol 313 ◽  
pp. 106719
Author(s):  
Timothy J. Keller ◽  
Alexander J. Laut ◽  
Jagadishwar Sirigiri ◽  
Thorsten Maly
Keyword(s):  
Nmr Spectroscopy ◽  
High Resolution ◽  
Magnetic Fields ◽  
Dynamic Nuclear Polarization ◽  
Nuclear Polarization ◽  
Proton Nmr ◽  
Proton Nmr Spectroscopy

Low-Field, Time-Resolved Dynamic Nuclear Polarization with Field Cycling and High-Resolution NMR Detection

2009 ◽  
Vol 37 (1-4) ◽  
pp. 515-537 ◽  
Author(s):  
S. E. Korchak ◽  
A. S. Kiryutin ◽  
K. L. Ivanov ◽  
A. V. Yurkovskaya ◽  
Yu. A. Grishin ◽  
...  
Keyword(s):  
High Resolution ◽  
Dynamic Nuclear Polarization ◽  
Nuclear Polarization ◽  
Time Resolved ◽  
Low Field ◽  
High Resolution Nmr ◽  
Field Cycling
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