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Salima Bahri1, Adil Safeer2, Agnes Adler2

  • 1NMR Spectroscopy, Bijvoet Center for Biomolecular Research, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands. s.bahri@uu.nl.

Journal of Biomolecular NMR
|June 8, 2023
PubMed
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This study introduces new proton-detected solid-state NMR methods for studying flexible biomolecules. These techniques enable detailed analysis of mobile protein sidechains and polysaccharides using standard equipment.

Area of Science:

  • Solid-state Nuclear Magnetic Resonance (NMR) spectroscopy
  • Biomolecular structure and dynamics
  • Macromolecular interactions

Background:

  • Solid-state NMR has advanced to study complex biomolecules, including large protein assemblies and intact cells at atomic resolution.
  • Highly flexible biomolecular components in insoluble environments pose challenges for traditional solution NMR.
  • Current methods for studying flexible regimes in solid-state NMR often involve 13C-detected experiments, partial deuteration, or ultra-fast magic-angle spinning (MAS).

Purpose of the Study:

  • To explore novel proton-detected pulse schemes for solid-state NMR.
  • To investigate the utility of these schemes for studying flexible biomolecules, such as mobile protein sidechains and polysaccharides.
  • To demonstrate the application of these methods using standard fast-spinning MAS probes at high magnetic fields.
Keywords:
Fungal cell wallMagic Angle SpinningMicrotubulesProtein dynamicsTau

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Main Methods:

  • Development and application of proton-detected pulse schemes probing through-bond 13C-13C correlations.
  • Utilized 2D and 3D spectroscopy for broadband analysis of flexible systems.
  • Employed standard fast-spinning MAS probes at high and ultra-high magnetic fields.

Main Results:

  • Successfully studied mobile protein sidechains and polysaccharides using the developed proton-detected pulse schemes.
  • Demonstrated the method's effectiveness in analyzing a mixture of microtubule-associated protein (MAP) tau and human microtubules (MTs).
  • Showcased the application to the cell wall of the fungus Schizophyllum commune, obtaining unambiguous correlations.

Conclusions:

  • The developed proton-detected pulse schemes are viable for studying flexible biomolecules and polysaccharides.
  • These methods provide unambiguous correlations, overcoming limitations of existing techniques.
  • The approach is applicable using standard fast-spinning MAS probes, enhancing accessibility for solid-state NMR research.