Creation of the analysis technique to visualize the dynamics of labile intermediates in phosphotransfer reactions
| Project/Area Number |
15K07887
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Physical pharmacy
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| Research Institution | Hiroshima University |
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Principal Investigator |
Kinoshita Emiko 広島大学, 医歯薬保健学研究科(薬), 助教 (40379912)
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | phos-tag / チオリン酸化タンパク質 / ATPgammaS / リン酸化タンパク質 / Phos-tag / リン酸基 / チオリン酸基 / ATPgS / リン酸基転移反応 / 反応中間体 / Phos-tag SDS-PAGE / ATPγS |
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| Outline of Final Research Achievements |
Signal-transduction regulatory systems for certain cellular responses which rely on phosphorylation of histidine or aspartic acid have known in eukaryotic cells. Identification of site-specific phosphorylation dynamics of His or Asp is technically difficult because of the labile nature of the phosphorylated amino acid residues. ATPgammaS, which used as a phosphoryl donor to trace protein kinase activities, has the advantage of permitting study of His- or Asp phosphorylation, because the resulting thiophosphorylated substrates are resistant to chemical hydrolysis or the action of phosphatases. Using the phosphate affinity electrophoresis, Phos-tag SDS-PAGE, we separated thiophosphorylated substrate generated as a intermediate of kinase reaction，and studied the dynamics of the histidine kinase reactions.
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Report
(4 results)
Research Products
(48 results)
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[Journal Article] A novel thiol-affinity micropipette tip method using zinc(II)-cyclen-attached agarose beads for enrichment of cysteine-containing molecules.2016
Author(s)
Kusamoto, H., Shiba, A., Koretake, N., Fujioka, H., Hieda, Y., Kinoshita-Kikuta, E., Kinoshita, E., Koike, T.
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Journal Title
J. Chromatogr. B. Analyt. Technol. Biomed. Life. Sci.
Volume: 1031
Pages: 195-201
DOI
Related Report
Peer Reviewed / Open Access / Acknowledgement Compliant
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[Journal Article] Expression and phosphorylation state analysis of intracellular protein kinases using Multi- PK antibody and Phos- tag SDS- PAGE.2015
Author(s)
Sugiyama, Y., Katayama, S., Kameshita, I., Morisawa, K., Higuchi, T., Todaka, H., Eiji Kinoshita, Kinoshita- Kikuta, E., Koike, T., Taniguchi, T., Sakamoto, S.
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Journal Title
MethodsX
Volume: 2
Pages: 469-474
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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