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NMR spectroscopic assignment of backbone and side-chain protons in fully protonated proteins: microcrystals, sedimented assemblies, and amyloid fibrils

Journal Article


Abstract


  • We demonstrate sensitive detection of alpha protons of fully protonated proteins by solid-state NMR spectroscopy with 100-111 kHz magic-angle spinning (MAS). The excellent resolution in the Cα-Hα plane is demonstrated for 5 proteins, including microcrystals, a sedimented complex, a capsid and amyloid fibrils. A set of 3D spectra based on a Cα-Hα detection block was developed and applied for the sequence-specific backbone and aliphatic side-chain resonance assignment using only 500 μg of sample. These developments accelerate structural studies of biomolecular assemblies available in submilligram quantities without the need of protein deuteration.

Authors


  •   Stanek, Jan (external author)
  •   Andreas, Loren (external author)
  •   Jaudzems, Kristaps (external author)
  •   Cala, Diane (external author)
  •   Lalli, Daniela (external author)
  •   Bertarello, Andrea (external author)
  •   Schubeis, Tobias (external author)
  •   Akopjana, Inara (external author)
  •   Kotelovica, Svetlana (external author)
  •   Tars, Kaspars (external author)
  •   Pica, Andrea (external author)
  •   Leone, Serena (external author)
  •   Picone, D (external author)
  •   Xu, Zhi-Qiang
  •   Dixon, Nicholas E.
  •   Martinez, Denis (external author)
  •   Berbon, Melanie (external author)
  •   El Mammeri, Nadia (external author)
  •   Noubhani, Abdelmajid (external author)
  •   Saupe, Sven (external author)
  •   Habenstein, Birgit (external author)
  •   Loquet, Antoine (external author)
  •   Pintacuda, Guido (external author)

Publication Date


  • 2016

Citation


  • Stanek, J., Andreas, L. B., Jaudzems, K., Cala, D., Lalli, D., Bertarello, A., Schubeis, T., Akopjana, I., Kotelovica, S., Tars, K., Pica, A., Leone, S., Picone, D., Xu, Z., Dixon, N. E., Martinez, D., Berbon, M., El Mammeri, N., Noubhani, A., Saupe, S., Habenstein, B., Loquet, A. & Pintacuda, G. (2016). NMR spectroscopic assignment of backbone and side-chain protons in fully protonated proteins: microcrystals, sedimented assemblies, and amyloid fibrils. Angewandte Chemie International Edition, 55 (50), 15504-15509.

Scopus Eid


  • 2-s2.0-85000415039

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/4255

Number Of Pages


  • 5

Start Page


  • 15504

End Page


  • 15509

Volume


  • 55

Issue


  • 50

Abstract


  • We demonstrate sensitive detection of alpha protons of fully protonated proteins by solid-state NMR spectroscopy with 100-111 kHz magic-angle spinning (MAS). The excellent resolution in the Cα-Hα plane is demonstrated for 5 proteins, including microcrystals, a sedimented complex, a capsid and amyloid fibrils. A set of 3D spectra based on a Cα-Hα detection block was developed and applied for the sequence-specific backbone and aliphatic side-chain resonance assignment using only 500 μg of sample. These developments accelerate structural studies of biomolecular assemblies available in submilligram quantities without the need of protein deuteration.

Authors


  •   Stanek, Jan (external author)
  •   Andreas, Loren (external author)
  •   Jaudzems, Kristaps (external author)
  •   Cala, Diane (external author)
  •   Lalli, Daniela (external author)
  •   Bertarello, Andrea (external author)
  •   Schubeis, Tobias (external author)
  •   Akopjana, Inara (external author)
  •   Kotelovica, Svetlana (external author)
  •   Tars, Kaspars (external author)
  •   Pica, Andrea (external author)
  •   Leone, Serena (external author)
  •   Picone, D (external author)
  •   Xu, Zhi-Qiang
  •   Dixon, Nicholas E.
  •   Martinez, Denis (external author)
  •   Berbon, Melanie (external author)
  •   El Mammeri, Nadia (external author)
  •   Noubhani, Abdelmajid (external author)
  •   Saupe, Sven (external author)
  •   Habenstein, Birgit (external author)
  •   Loquet, Antoine (external author)
  •   Pintacuda, Guido (external author)

Publication Date


  • 2016

Citation


  • Stanek, J., Andreas, L. B., Jaudzems, K., Cala, D., Lalli, D., Bertarello, A., Schubeis, T., Akopjana, I., Kotelovica, S., Tars, K., Pica, A., Leone, S., Picone, D., Xu, Z., Dixon, N. E., Martinez, D., Berbon, M., El Mammeri, N., Noubhani, A., Saupe, S., Habenstein, B., Loquet, A. & Pintacuda, G. (2016). NMR spectroscopic assignment of backbone and side-chain protons in fully protonated proteins: microcrystals, sedimented assemblies, and amyloid fibrils. Angewandte Chemie International Edition, 55 (50), 15504-15509.

Scopus Eid


  • 2-s2.0-85000415039

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/4255

Number Of Pages


  • 5

Start Page


  • 15504

End Page


  • 15509

Volume


  • 55

Issue


  • 50