2012 Fiscal Year Final Research Report
Development of NMR measurement and analysis method for protein structural study in 2H2O solution.
| Project/Area Number |
22570109
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Structural biochemistry
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| Research Institution | Hokkaido University |
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Principal Investigator |
OGURA Kenji 北海道大学, 大学院・先端生命科学研究院, 特任准教授 (50270682)
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| Project Period (FY) |
2010 – 2012
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| Keywords | NMR / タンパク質 / 立体構造解析 |
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| Research Abstract |
We developed an NMR pulse sequence, 3D HCA(N)CO, to correlate the chemical shifts of protein backbone 1H and 13C to those of 13C' in the preceding residue. 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.
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