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A new capillary mounting technique for protein microcrystals and its derivatization by xenon

Research Project

Project/Area Number 23600016
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Quantum beam science
Research InstitutionJapan Synchrotron Radiation Research Institute

Principal Investigator

KUMASAKA Takashi  公益財団法人高輝度光科学研究センター, タンパク質結晶解析推進室, 副主席研究員 (30291066)

Co-Investigator(Renkei-kenkyūsha) MAKINO Masatomo  島根県産業技術センター, 研究員 (30529582)
Project Period (FY) 2011-04-28 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2011: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywords結晶解析 / タンパク質 / 結晶構造解析 / 希ガス / 微小結晶 / 試料処理 / 機器分析 / 蛋白質
Outline of Final Research Achievements

Synchrotron microbeam technology has opened the new window of protein crystallography―enabling us to analyze atomic structures from the microcrystals with the size of ca. 10 um. Such tiny and fragile crystals, however, were difficult to be handled in 2011 when this project has been started. To solve the issue, we developed a mounting technique inspired by a capillary used for microinjection in cell biology.
Glass capillary is rigid, therefore it can be made sharper. The fine and tapered capillary, with 15 um diameter at its tip, successfully held and cooled the crystals. The capillary can minimize amount of solvent, leading to the reduction of X-ray scattering noise.
Since the capillary is resistant to pressure, it is also used as a gas-derivatization tool. The capillary attached to a screw-type mounting tool can directly connect to the gas-pressurization device. By a simple procedure, the xenon-derivatives of egg-white lysozyme crystals was prepared and phased by X-ray anomalous data.

Report

(5 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • 2013 Research-status Report
  • 2012 Research-status Report
  • 2011 Research-status Report
  • Research Products

    (6 results)

All 2013 2012 Other

All Journal Article (2 results) (of which Peer Reviewed: 2 results) Presentation (2 results) Book (1 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] A convenient tool for gas derivatization using fine-needle capillary mounting for protein crystals.2013

    • Author(s)
      Mizuno, N., Makino, M., Kumasaka, T.
    • Journal Title

      J. Sync. Rad.

      Volume: 20 Issue: 6 Pages: 999-1002

    • DOI

      10.1107/s0909049513021584

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Fine-needle capillary mounting for protein microcrystals.2012

    • Author(s)
      M. Makino, I. Wada, N. Mizuno, K. Hirata, N. Shimizu, T. Hikima, M. Yamamoto and T. Kumasaka
    • Journal Title

      J. Appl. Cryst.

      Volume: 45 Issue: 4 Pages: 785-788

    • DOI

      10.1107/s0021889812024545

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Presentation] Fine-needleキャピラリーを用いたタンパク質結晶マウント法2012

    • Author(s)
      水野伸宏、牧野正知、和田いづみ、平田邦生、引間孝明、山本雅貴、熊坂崇
    • Organizer
      平成24年度日本結晶学会年会
    • Place of Presentation
      東北大学(宮城県仙台市)
    • Related Report
      2012 Research-status Report
  • [Presentation] Advanced Crystal Mounting Tool for Gas Pressurization Improves Efficiency of Xenon-Derivatization

    • Author(s)
      Nobuhiro Mizuno, Masatomo Makino, Takashi Kumasaka
    • Organizer
      The 12th Conference of the Asia Crystallographic Association
    • Place of Presentation
      HKUST, Hong Kong
    • Related Report
      2013 Research-status Report
  • [Book] Fine-needleキャピラリーを用いたマウント法(「タンパク質結晶の最前線」所収)2013

    • Author(s)
      水野伸宏, 熊坂崇
    • Total Pages
      9
    • Publisher
      シーエムシー出版
    • Related Report
      2013 Research-status Report
  • [Patent(Industrial Property Rights)] 微小試料用キャピラリー2012

    • Inventor(s)
      熊坂崇,牧野正知,桑本いづみ,山本雅貴
    • Industrial Property Rights Holder
      熊坂崇,牧野正知,桑本いづみ,山本雅貴
    • Industrial Property Number
      2012-018603
    • Filing Date
      2012
    • Related Report
      2011 Research-status Report

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Published: 2011-08-05   Modified: 2019-07-29  

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