2002 Fiscal Year Final Research Report Summary
Development of the ^<31>P-NMR Probe with ultra high sensitivity and analysis of phosphate binding proteins, related to signal transduction
| Project/Area Number |
13558080
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Structural biochemistry
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| Research Institution | THE UNIVERSITY OF TOKYO |
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Principal Investigator |
TANOKURA Masaru University of Tokyo, Graduate School of Agricultural and sciences, Professor, 大学院・農学生命科学研究科, 教授 (60136786)
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| Co-Investigator(Kenkyū-buntansha) |
HARUYAMA Hideki Sankyo Co., Ltd., Biomedical Research Laboratories, Vice-chief of laboratory (Reseacher), バイオメディカル研究所, 所次長(研究職)
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| Project Period (FY) |
2001 – 2002
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| Keywords | ^<31>P NMR / Probe / Phosphate binding protein |
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| Research Abstract |
Although the sensitivity of ^<31>P-NMR (nuclear magnetic resonance) is a comparatively high and phosphate is existed mostly in forms, such as ATP, in the living body, ^<31>P is difficult for obtaining useful data because T_2 and T_1 are very short and long, respectively. Therefore, the sensitivity of the NMR measurement obtained so far has been a serious obstacle when advancing a research for performing an experiment to detect the signals from a sample containing single cells. However, the strength of the magnetic field used for NMR measurement goes up and probe technology was also improved in recent years, a possibility of measuring and analyzing by ^<31>P-NMR the single cells has also increased.
We have developed the probe equipment and the cell for detecting can carry out a direct monitoring the single cells in a condition making physiological environment. This probe can be improving a pulse for measurement and a receiver's dynamic range so that it can respond to short T_2 especially. Thus equipment hold the cell and the tissue in the state where it lived for a long time, it made it possible to measure the ^<31>P-NMR spectra. Moreover, it analyzed also about the milk, which is the mixture of the living substances used as the candidate for detection in this probe. It succeeded in assigning the phosphate compound as an ingredient in milk by applying a 2D ^1H-^<31>P NMR spectra in addition to ^1D ^<31>P NMR measurement, without pretreating.
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[Publications] Sakai, N., Yao, M., Itou, H., Watanabe, N., Yumoto, F., Tanokura, M., Tanaka, I.: "The three-dimensional stricture of septum site-determining protein MinD from Pyrococcus horikoshii OT3 in complex with Mg-ADP."Structure. 9. 817-826 (2001)
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「研究成果報告書概要(欧文)」より
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[Publications] Yumoto, F., Nara, M., Kagi, H., Iwasaki, W., Ojima, T., Nishita, K., Nagata, K., Tanokura, M.: "Coordination strictures of Ca^<2+>+ and Mg^<2+>+ in Akazara scallop troponin C in solution : FTIR spectroscopy of side-chain COO^-groups."Eur.J.Biochem.. 268. 6284-6290 (2001)
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[Publications] Iwasaki, W., Sasaki, H., Nakamura, A., Kohama K., Tanokura, M.: "Metal-free and Ca^<2+>-bound strictures of a multidomain EF-hand protein, CBP40, from the lower eukaryote, Physarum polycephalum."Structure. 11. 75-85 (2003)
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「研究成果報告書概要(欧文)」より
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