• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Construction of capillary electrophoresis/mass spectrometry interface

Research Project

Project/Area Number 07558219
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Structural biochemistry
Research InstitutionFujita Health University

Principal Investigator

TITANI Koiti  Inst.For Comprehensive Med.Sci., Fujita Health University, Professor, 総合医科学研究所, 教授 (60179942)

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)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 1997: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1996: ¥2,700,000 (Direct Cost: ¥2,700,000)
Keywordsmass spectrometry / capillary electrophoresis / protein / nucleic acid / structural analysis / 電気泳動 / 蛋白質
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.

Report

(4 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • Research Products

    (39 results)

All Other

All Publications (39 results)

  • [Publications] Morishima-Kawashima, M.et al.: "Proline-Directed and Non-Proline-Directed Phosphorylation of PHF-tau." J.Biol.Chem.270. 823-829 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kanamori, T.et al.: "Identification of Two 17 kDa Rat Parotid Gland Phosphoproteins,Subjects for Dephosphorylation upon β-Adrenergic Stimulation, as Destrin-and Cofilin-like Proteins." J.Biol.Chem.270. 8061-8067 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Morishima-Kawashima, M., et al.: "Hyperphosphorylation of Tau in PHF" Neurobiol.Aging. 16. 365-380 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Koike, T., et al.: "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)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hayashi, N., et al.: "Structural Studies of tRNAs by Capillary High Pertormance Liquid Chromatography/Electrospray Mass Spectrometry." Nucleic Acids Symp. Ser.34. 153-154 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Usami, Y., et al.: "Primary Structure of Alboaggregin-B Purified from the Venom of Trimeresurus albolabris." Biochem. Biophys. Res. Commun.219. 727-733 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 谷口 寿章: "キャピラリーLC/MS法による蛋白質・核酸の修飾の解析" J.Mass Spectrom.Soc. Japan. 44. 443-457 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Matsubara M., et al.: "Site-specific Phosphorylation of Synapsin I by Motogenactivated Prorein Kinase and Cdk5 and Its Effects on Physiological Functions." J.Biol, Chem. 271. 21108-21113 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hayashi N.et al.: "Circular dichroism and ^1H nuclear magnetic resonance, studies on the solution and membrane structures of GAP-43 calmodulin-binding domain." J.Biol. Chem.272. 7639-7645 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ymauchi, E., et sl.: "The C-terminal conserved domain of marcks is phosphorlatde in vivo by proline-directed protein kinase:application of ion trap mass spectrometry to the determination of protein phosphoreylation sites." J.Biol. Chem.273. 4367-4371 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Taniguchi, H., and Hayashi, N.: "A liquid chromatography/electrospray mass spectrometric study on the posttranscriptional modification of tRNA." Nucleic Acids Res.26. 1481-1486 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Yamauchi, E., et al.: "Presence of conserved domains in the C-terminus of MARCKS, a major in vivo substrate of protein kinase C:application of ion trap mass spectrometry to the elucidation of protein structures" J.Biochem.(印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [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)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kanamori, T., Hayakawa, T., Suzuki, M., and Titani K.: "Identification of Two 17 kDa Rat Parotid Gland Phosphoproteins, Subjects for Dephosphorylationupon beta-Adrenergic Stimulation, as Destrin-and Cofilin-like Proteins." J.Biol.Chem.270. 8061-8067 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [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)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [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)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hayashi, N., Suzuki, M., Titani, K., and Taniguchi, H.: "Structural Studies of tRNAs by Capillary High Performance Liquid Chromatography/Electrospray Mass Spectrometry." Nucleic Acids Symp.Ser.34. 153-154 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [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)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [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)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hayashi, N., Matsubara, M., Titani, K., and Taniguchi, H.: "Circular dichroism and ^1H nuclear magnetic resonance, studies on the solution and membrane structures of GAP-43 calmodulin-binding domain." J.Biol.Chem.272. 7639-7645 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Yamauchi, E., Kiyonami, R., Kanai, M., and Taniguchi, H.: "The C-terminal is phosphorylated in vivo by proline-directed protein kinase : application of ion trap mass spectrometry to the determination of protein phosphorylation sites." J.Biol.Chem.273. 4367-4371 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Taniguchi, H., and Hayashi, N.: "A liquid chromatography/electrospray mass spectrometric study on the posttranscriptional modification of tRNA." Nucleic Acids Res.26. 1481-1486 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Yamauchi, E., Kiyonami, R., Kanai, M., and Taniguchi, H.: "Presence of conserved domains in the C-terminus of MARCKS,a major in vivo substrate of protein kinase C : application of ion trap mass spectrometry to the elucidation of protein structures" J.Biochem.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Yamauchi,E.,et al.: "The C-terminal conserved domain of marcks is phosphorylated in vivo by proline-directed protein linase: application of ion trap mass spectrometry to the determination of protein phosphorylation sites." J.Biol.Chem.273. 4367-4371 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Takeda,S.,et al.: "Structural and functional domains of the troponin complex revealed by limited digestion." Eur.J.Biochem.246. 611-617 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Saijo,Y.,et al.: "Production, cryatallization and preliminary X-ray analysis of rabbit skeletal muscle troponin complex consisting of troponin C and fragment (1-47) of troponin I" Protein Sci.6. 916-918 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Hayashi,N.,et al.: "Circular dichroism and ^1H nuclear magnetic resonance studies on the solution and membrane structures of Gap-43 calmodulin-binding domain." J.Biol.Chem.272. 7639-7645 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Taniguchi,H.,and Hayashi,N.: "A liquid chromatography/electrospray mass spectrometric study on the posttranscriptional modification of tRNA." Nucleic Acids Res.26. 1481-1486 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] 谷口寿章: "キャピラリーLC/MS法による蛋白質・核酸の修飾の解析" J.Mass Spectron.Soc.Japan. 44. 443-457 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] M.Matsubara,et al.: "Site-specific Phosphorylation of Synapsin I by Motogen-activated Prorein Kinase and Cdk5 and Its Effects on Physiological Functions." J.Biol.Chem.271. 21108-21113 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Morishima-Kawashima, M. et al.: "Proline-Directed and Non-Proline-Directed Phosphorylation of PHF-tau." J. Biol. Chem.270. 823-829 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kanamori, T., et al.: "Identification of Two 17 KDa Rat Parotid Gland Phosphoproteins, Subjects for Dephosphorylation upon β-Adrenergic Stimulation, as Destrin-and Cofilin-like Proteins." J. Biol. Chem.270. 8061-8067 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Morishima-Kawashima, M., et al.: "Hyperphosphorylation of Tau in PHF." Neurobiol. Aging. 16. 365-380 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Koike, T., et al.: "The Complete Amino Acid Sequence of a Mannose-binding Lectin from “Kidachi Aloe" (Aloe arborescens Miller ver. natalensis Berger)." Biochem. Biophys. Res. Commun.214. 163-170 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Hayashi, N., et al.: "Structural Studies of tRNAs by Capillary High Performance Liquid Chromatography/ Electrospray Mass Spectrometry." Nucleic Acids Symp. Ser.34. 153-154 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Usami, Y., et al.: "Primary Structure of Alboagggregin-B Purified from the Venom of Trimeresurus albolabris." Biochem. Biophys. Res. Commun.(in press). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kawasaki, T., et al.: "Complete Amino Acid Sequence and Indentification of the Platelet GPIb Binding Site of Jararaca GPIb-BP, a Snake Venom Protein Isolated from Bothrops jararaca." J. Biol. Chem.(in press). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Manenti, S. et al.: "Demyristoylation of Myristoylated Alanine-Rich C Kinase Substrate." Biochem. Soc. Trans.23. 561-564 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 谷口 寿章: "キャピラリーLC/MS法による蛋白質・核酸の修飾の解析" J. Mass Spectrom. Soc. Japan. (印刷中). (1996)

    • Related Report
      1995 Annual Research Report

URL: 

Published: 1996-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi