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Facilitation of Crystallization of Proteins by Trimming

Research Project

Project/Area Number 03454564
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 生物物性学
Research InstitutionHYOGO UNIVERSITY OF TEACHER EDUCATION

Principal Investigator

SHIRAKIHARA Yasuo  Hyogo University of Teacher Education, Faculty of School Education Associate Professor, 学校教育学部, 助教授 (20150287)

Project Period (FY) 1991 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1992: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1991: ¥5,400,000 (Direct Cost: ¥5,400,000)
KeywordsX-ray crystallography / Three-dimensional Structure of Proteins / Crystallization / Proteolytic Enzyme
Research Abstract

In order to accelerate to get the structure of the proteins by x-ray protein crystallography, a trial has been made to facilitate crystal growth by trimming the target protein appropriately. This trial is in accordance with calls for more independent 3-dimensional structure information.
The target proteins were:F1-ATPase from a thermophilic bacterium, its alpha3beta3 complex, rat DNA polymerase beta, Env Z protein, PhoB protein, Band 3 protein, CamR repressor, Asp racemase and flagellin. Those proteins were subjected to a limited proteolysis by a number of proteases to look for appropriately trimmed molecules. The trimmed molecules were then purified and subjected to crystallization experiment.
The results are: 1.In case of rat DNA polymerase beta, a large C-terminal fragment (representing 80 % of the molecule) was found to crystallize to a suitable form. The intact molecule resisted to crystallization. The crystals diffracted to at least to 2.5 A^^゚ resolution, and the data collection and derivative search is now in progress. This indicates that the trimmed molecule does form good crystals. 2.41 k fragment of flagellin formed crystals. The intact flagellin formed helical aggregates under most of crystallization conditions. Although crystals were formed, the conditions are to be refined to get better crystals. 3.Appropriate fragments of PhoB protein and CamR repressor were found. They are now subjected to purification.

Report

(3 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • Research Products

    (4 results)

All Other

All Publications (4 results)

  • [Publications] 白木原 康雄,松影 昭夫,西本 義男,伊達 孝保: "ラットDNAポリメラーゼβの結晶-トリプシン処理の効用-" 日本生物物理学会「生物物理」. 32. 5119 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Yasuo Shirakihara, Akio Matsukage, Yoshio Nishimoto & Masayasu Date: "Crystallization of Rat DNA polymerase beta-Effect of Trypsin Treatment-" Biophysics. 32. S119 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 白木原 康雄,松影 昭夫 西本 義男,伊達 孝保: "ラットDNAポリメラーゼβの結晶化ートリプシン処理の効用ー" 日本生物物理学会「生物物理」. 32. S119- (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 白木原 康雄,松影 昭夫: "分子整形を行なったラットDNAポリメラ-ゼβの結晶化" 日本生物物理学会年会講演予稿集(口頭発表).

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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