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Protein residue linking in a single spectrum for magic-angle spinning NMR assignment

Journal Article


Abstract


  • Here we introduce a new pulse sequence for resonance assignment that halves the number of data sets required for sequential linking by directly correlating sequential amide resonances in a single diagonal-free spectrum. The method is demonstrated with both microcrystalline and sedimented deuterated proteins spinning at 60 and 111 kHz, and a fully protonated microcrystalline protein spinning at 111 kHz, with as little as 0.5 mg protein sample. We find that amide signals have a low chance of ambiguous linkage, which is further improved by linking in both forward and backward directions. The spectra obtained are amenable to automated resonance assignment using general-purpose software such as UNIO-MATCH.

Authors


  •   Andreas, Loren (external author)
  •   Stanek, Jan (external author)
  •   Le Marchand, Tanguy (external author)
  •   Bertarello, Andrea (external author)
  •   Paepe, Diane C-D. (external author)
  •   Lalli, Daniela (external author)
  •   Krej¿íková, Magdaléna (external author)
  •   Doyen, Camille (external author)
  •   Ã–ster, Carl (external author)
  •   Knott, Benno (external author)
  •   Wegner, Sebastian (external author)
  •   Engelke, Frank (external author)
  •   Felli, Isabella C. (external author)
  •   Pierattelli, Roberta (external author)
  •   Dixon, Nicholas E.
  •   Emsley, Lyndon (external author)
  •   Herrmann, Torsten (external author)
  •   Pintacuda, Guido (external author)

Publication Date


  • 2015

Citation


  • Andreas, L. B., Stanek, J., Le Marchand, T., Bertarello, A., Paepe, D. Cala-De., Lalli, D., Krejcikova, M., Doyen, C., Öster, C., Knott, B., Wegner, S., Engelke, F., Felli, I. C., Pierattelli, R., Dixon, N. E., Emsley, L., Herrmann, T. & Pintacuda, G. (2015). Protein residue linking in a single spectrum for magic-angle spinning NMR assignment. Journal of Biomolecular NMR, 62 (3), 253-261.

Scopus Eid


  • 2-s2.0-84944441197

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/3005

Has Global Citation Frequency


Number Of Pages


  • 8

Start Page


  • 253

End Page


  • 261

Volume


  • 62

Issue


  • 3

Place Of Publication


  • Netherlands

Abstract


  • Here we introduce a new pulse sequence for resonance assignment that halves the number of data sets required for sequential linking by directly correlating sequential amide resonances in a single diagonal-free spectrum. The method is demonstrated with both microcrystalline and sedimented deuterated proteins spinning at 60 and 111 kHz, and a fully protonated microcrystalline protein spinning at 111 kHz, with as little as 0.5 mg protein sample. We find that amide signals have a low chance of ambiguous linkage, which is further improved by linking in both forward and backward directions. The spectra obtained are amenable to automated resonance assignment using general-purpose software such as UNIO-MATCH.

Authors


  •   Andreas, Loren (external author)
  •   Stanek, Jan (external author)
  •   Le Marchand, Tanguy (external author)
  •   Bertarello, Andrea (external author)
  •   Paepe, Diane C-D. (external author)
  •   Lalli, Daniela (external author)
  •   Krej¿íková, Magdaléna (external author)
  •   Doyen, Camille (external author)
  •   Ã–ster, Carl (external author)
  •   Knott, Benno (external author)
  •   Wegner, Sebastian (external author)
  •   Engelke, Frank (external author)
  •   Felli, Isabella C. (external author)
  •   Pierattelli, Roberta (external author)
  •   Dixon, Nicholas E.
  •   Emsley, Lyndon (external author)
  •   Herrmann, Torsten (external author)
  •   Pintacuda, Guido (external author)

Publication Date


  • 2015

Citation


  • Andreas, L. B., Stanek, J., Le Marchand, T., Bertarello, A., Paepe, D. Cala-De., Lalli, D., Krejcikova, M., Doyen, C., Öster, C., Knott, B., Wegner, S., Engelke, F., Felli, I. C., Pierattelli, R., Dixon, N. E., Emsley, L., Herrmann, T. & Pintacuda, G. (2015). Protein residue linking in a single spectrum for magic-angle spinning NMR assignment. Journal of Biomolecular NMR, 62 (3), 253-261.

Scopus Eid


  • 2-s2.0-84944441197

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/3005

Has Global Citation Frequency


Number Of Pages


  • 8

Start Page


  • 253

End Page


  • 261

Volume


  • 62

Issue


  • 3

Place Of Publication


  • Netherlands