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Chemical Shift: Internal References and Solvent Effects01:17

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In an NMR sample, precise measurement of the absolute absorption frequencies of nuclei is difficult. A standard internal reference compound is added, and the frequency difference between the reference signal and sample signals is measured.
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The spin state of an NMR-active nucleus can have a slight effect on its immediate electronic environment. This effect propagates through the intervening bonds and affects the electronic environments of NMR-active nuclei up to three bonds away; occasionally, even farther. This phenomenon is called spin–spin coupling or J-coupling. Coupling interactions are mutual and result in small changes in the absorption frequencies of both nuclei involved. While nuclei of the same element are involved...
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Aromatic compounds can be identified or analyzed using proton NMR and carbon‐13 NMR. Typically, aromatic hydrogens or hydrogens directly bonded to the aromatic rings are strongly deshielded by the aromatic ring current. Therefore, they absorb in the range of 6.5–8.0 ppm in proton NMR spectra. For instance, aromatic hydrogens directly bonded to the benzene ring absorb at 7.3 ppm. However, aromatic hydrogens of larger rings absorb farther upfield or downfield than the ideal range.
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Method to Visualize and Analyze Membrane Interacting Proteins by Transmission Electron Microscopy
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Membrane-embedded TSPO: an NMR view.

Gwladys Rivière1,2, Garima Jaipuria1,2, Loren B Andreas3

  • 1Senior Research Group of Translational Structural Biology in Dementia, Deutsches Zentrum für Neurodegenerative Erkrankungen (DZNE), Von-Siebold-Str. 3a, 37075, Göttingen, Germany.

European Biophysics Journal : EBJ
|December 23, 2020
PubMed
Summary
This summary is machine-generated.

The structure of Translocator Protein (18 kDa) (TSPO) is influenced by its lipid environment. Cholesterol specifically impacts TSPO

Keywords:
CholesterolDynamicsLipidMembrane proteinNeurodegenerationSolid-state NMRStructureTSPO

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Area of Science:

  • Biochemistry
  • Structural Biology
  • Neuroscience

Background:

  • Translocator Protein (18 kDa) (TSPO) is a key mitochondrial protein implicated in neuroinflammation.
  • TSPO serves as a biomarker for neurodegenerative diseases and a potential therapeutic target.
  • The precise function of TSPO remains largely unknown, hindering therapeutic development.

Purpose of the Study:

  • To investigate the influence of lipid composition on the structure of mammalian TSPO.
  • To elucidate the role of cholesterol in TSPO's tertiary, quaternary, and localization dynamics.
  • To provide insights into TSPO's cellular activities for improved ligand design.

Main Methods:

  • Solid-state Nuclear Magnetic Resonance (ssNMR) experiments were employed.
  • Mammalian TSPO was embedded within lipid bilayers to mimic native environments.
  • Structural analysis focused on the impact of varying lipid compositions, particularly cholesterol.

Main Results:

  • Lipid composition significantly affects the structure of TSPO within lipid bilayers.
  • Cholesterol was identified as a key modulator of both tertiary and quaternary TSPO structures.
  • Cholesterol influences the localization of TSPO within mitochondria-associated endoplasmic reticulum membranes.

Conclusions:

  • Understanding TSPO's lipid-dependent structure is crucial for deciphering its function.
  • Cholesterol plays a significant role in regulating TSPO's structural conformation and cellular positioning.
  • These findings are essential for developing targeted diagnostics and therapeutics for neurodegenerative diseases.