1997 Fiscal Year Final Research Report Summary
Construction of capillary electrophoresis/mass spectrometry interface
| Project/Area Number |
07558219
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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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 | Fujita Health University |
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Principal Investigator |
TITANI Koiti Inst.For Comprehensive Med.Sci., Fujita Health University, Professor, 総合医科学研究所, 教授 (60179942)
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| Co-Investigator(Kenkyū-buntansha) |
TSUNASAWA Susumu Research Institute, Takara Shuzo Co.Vice Director, バイオ研究所, 副所長
SUZUKI Masami Inst.For Comprehensive Med.Sci., Fujita Health University, Research associate, 総合医科学研究所, 研究員
TANIGUCHI Hisaaki Inst.For Comprehensive Med.Sci., Fujita Health University, Assist.Professor, 総合医科学研究所, 助教授 (10257636)
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| Project Period (FY) |
1995 – 1997
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| Keywords | mass spectrometry / capillary electrophoresis / protein / nucleic acid / structural analysis |
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| Research Abstract |
In the first fiscal year, we made a coaxial type capillary electrophoresis/mass spectrometry (CE/MS) interface for the connection between the existing electrospray mass spectrometer (Sciex API III) and the newly purchased capillary electroresis (CE) apparatus (Beckman P/ACE 5000). During the second year, the efforts were concentrated in the optimization of the electrophoresis conditions that are compatible with the electrospray ionization. The results obtained were very close to our initial expectations ; the resolution and the sensitivity observed with the new interface were superior to those obtained with the existing liquid-junction type interface. Furthermore, the amounts of samples needed for the analysis are minimal. The amounts are less than one percent of those used for the previous LC/MS analysis. This is a clear advantage in the protein analysis, since the rest of the samples can be used to other analyzes such as amino acid sequencing. In the third (final) year, we introduced the so-called nanospray ionization method to achieve higher sensitivities. For this purpose, we established the existing nanospray method using glass microinjection capillary as well as the new microspray method using pulled-fused silica capillaries. The latter method allowed us to get stable electrospray ionization at a flow rate around 20-50 nl/min. This method, therefore, can be well combined with the capillary electrophoresis.
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[Publications] Morishima-Kawashima, M., Hasegawa, M., Takio, K., Suzuki, M., Yoshida, H., Titani, K., and Ihara, Y.: "Proline-Directed and Non-Proline-Directed Phosphorylation of PHF-tau." J.Biol.Chem.270. 823-829 (1995)
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「研究成果報告書概要(欧文)」より
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[Publications] Morishima-Kawashima, M., Hasegawa, M., Takio, K., Suzuki, M., Yoshida, H., Watanabe, A., Titani, K., and Ihara, Y.: "Hyperphosphorylation of Tau in PHF." Neurobiol.Aging. 16. 365-380 (1995)
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[Publications] Koike, T., Titani, K., Suzuki, M., Beppu, H., Kuzuya, H., Maruta, K., Shimpo, K., and Fujita, K.: "The Complete Amino Acid Sequence of a Mannose-binding Lectin from "Kidachi Aloe" (Aloe arborescens Miller var.natalensis Berger)." Biochem.Biophys.Res.Commun.214. 163-170 (1995)
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[Publications] Usami, Y., Suzuki, M., Yoshida, E., Sakurai, Y., Hirano, K., Kawasaki, T., Fujimura, Y., and Titani, K.: "Primary Structure of Alboagggregin-B Purified from the Venom of Trimeresurus albolabris." Biochem.Biophys.Res.Commun.219. 727-733 (1996)
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[Publications] Matsubara M., Kusubata, M., Ishiguro, K., Uchida, T., Titani, K., and Taniguchi, H.: "Site-specific Phosphorylation of Synapsin I by Motogen-activated Prorein Kinase and Cdk5 and Its Effects on Physiological" J.Biol.Chem.271. 21108-21113 (1996)
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