Structure determination of proteins in 2H2O solution aided by a deuterium-decoupled 3

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Abstract We developed an NMR pulse sequence, 3D HCA(N)CO, to correlate the chemical shifts of protein backbone 1Hα and 13Cα to those of 13C� in the preceding residue. By applying 2H decoupling, the experiment was accomplished with high sensitivity comparable to that of HCA(CO)N. When combined with HCACO, HCAN and HCA(CO)N, the HCA(N)CO sequence allows the sequential assignment using backbone 13C� and amide 15N chemical shifts without resort to backbone amide protons. This assignment strategy was demonstrated for 13C/15N-labeled GB1 dissolved in 2H2O. The quality of the GB1 structure determined in 2H2O was similar to that determined in H2O in spite of significantly smaller number of NOE correlations. Thus this strategy enables the determination of protein structures in 2H2O or H2O at high pH values.

• Content Type Journal Article
• DOI 10.1007/s10858-010-9431-y
• Authors
  • Kenji Ogura, Hokkaido University Department of Structural Biology, Faculty of Advanced Life Science Kita 21 Nishi 11, Kita-ku Sapporo 001-0021 Japan
  • Hiroyuki Kumeta, Hokkaido University Department of Structural Biology, Faculty of Advanced Life Science Kita 21 Nishi 11, Kita-ku Sapporo 001-0021 Japan
  • Fuyuhiko Inagaki, Hokkaido University Department of Structural Biology, Faculty of Advanced Life Science Kita 21 Nishi 11, Kita-ku Sapporo 001-0021 Japan

• • Journal Journal of Biomolecular NMR
  • Online ISSN 1573-5001
  • Print ISSN 0925-2738
  • Journal Volume Volume 47
  • Journal Issue Volume 47, Number 4

Source: Journal of Biomolecular NMR