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Studies on defect generation mechanisms in protein crystals by molecular-level in situ observations of elementary growth processes

Research Project

Project/Area Number 16360001
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied materials science/Crystal engineering
Research InstitutionTohoku University

Principal Investigator

SAZAKI Gen  Tohoku University, Institute for Materials Research, Lecturer, 金属材料研究所, 講師 (60261509)

Co-Investigator(Kenkyū-buntansha) TSUKAMOTO Katsuo  Tohoku University, Faculty of Science, Associate Professor, 大学院・理学研究科, 助教授 (60125614)
HIGUCHI Hideo  Tohoku University, Biomedical Engineering Research Organization, Professor, 先進医工学研究機構, 教授 (90165093)
Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥15,400,000 (Direct Cost: ¥15,400,000)
Fiscal Year 2005: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 2004: ¥13,100,000 (Direct Cost: ¥13,100,000)
KeywordsProtein / Crystal growth / Lattice defects / In situ observation / Elementary growth steps / Surface diffusion / Laser confocal microscopy / Single molecule visualization / レーザー共焦点顕微鏡
Research Abstract

Using tetragonal crystals of model protein lysozyme, we observed in situ (i) elementary growth steps on a crystal surface, (ii) individual protein molecules diffusing at a crystal-solution interface and (iii) defects inside a crystal by advanced optical microscopy : laser confocal microscopy combined with differential interference contrast microscopy (LCM-DIM), a single molecule visualization technique (SMV) and laser light scattering tomography (LLST). Key results found in this study were as follows.
1) Effects of flow : In situ observation of growth steps on a crystal surface under forced flow by LCM-DIM enable us to find that flow significantly accelerates the transport of impurity to a crystal surface, and then provide bunched steps.
2) Surface diffusion : We succeeded in the in situ observation of individual fluorescent-labeled lysozyme (F-L) molecules diffusing in the vicinity of a crystal surface by SMV, for the first time. We found that diffusion of F-L molecules at a crystal-sol … More ution interface is 4 orders of magnitude slower than that in a bulk solution, and anisotropy of diffusion exists according to crystallographic directions.
3) Adsorption mechanism of impure protein : From in situ observation of adsorption processes of F-L and fluorescent-labeled dimmer of lysozyme (F-D) on a crystal surface by both LCM-DIM and SMV simultaneously, we demonstrated that F-L adsorbs preferentially on a step, whereas F-D adsorbs randomly on a terrace. The different adsorption sites of F-L and F-D provided different supersaturation dependencies of impurity effects.
4) Defects inside crystals : We succeeded in observing in situ dislocations and inclusions inside a growing crystal by LCM-DIM, and revealed that microcrystals incorporated in a seed crystal generate many dislocations and hence provide spiral growth hillocks on top of microcrystals incorporated. We also succeeded in visualizing the contrasts that are probably microdefects originated from vacancies and impurity particles inside a crystal by LLST. Less

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • Research Products

    (39 results)

All 2006 2005 2004 Other

All Journal Article (32 results) Book (6 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] Structural consequences of hen egg-white lysozyme orthorhombic crystal growth in a high magnetic field : validation of X-ray diffraction intensity, conformational energy searching and quantitative analysis of B factors and mosaicity2005

    • Author(s)
      S.Saijo
    • Journal Title

      Acta Cryst. D61

      Pages: 207-217

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Kinetic acceleration of the growth rate of glucose isomerase crystals under high pressure2005

    • Author(s)
      Y.Suzuki
    • Journal Title

      J. Phys. Chem. B109

      Pages: 3222-3226

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Investigations on electromigration phenomena for protein crystallization using crystal growth cells with multiple electrode : Effect of the potential control2005

    • Author(s)
      E.Nieto-Mendoza
    • Journal Title

      J. Crystal Growth 275

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] In situ observation of dislocations in protein crystals during growth by advanced optical microscopy2005

    • Author(s)
      G.Sazaki
    • Journal Title

      Crystal Growth and Design 5

      Pages: 1729-1735

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] 格子欠陥発生機構の解明に向けたタンパク質結晶表面の高分解光学観察2005

    • Author(s)
      佐崎 元
    • Journal Title

      応用物理 74

      Pages: 608-612

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Kinetic acceleration of the growth rate of glucose isomerase crystals under high pressure2005

    • Author(s)
      Y.Suzuki
    • Journal Title

      J.Phys.Chem.B 109

      Pages: 3222-3226

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Investigations on electromigration phenomena for protein crystallization using crystal growth cells with multiple electrode : Effect of the potential control2005

    • Author(s)
      E.Nieto-Mendoza
    • Journal Title

      J.Crystal Growth 275

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Surface observation of protein crystals by high-resolution optical microscopy2005

    • Author(s)
      G.Sazaki
    • Journal Title

      Oyo Butsuri 74

      Pages: 608-612

    • NAID

      10015722624

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] In situ observation of dislocations in protein crystals during growt by advanced optical microscopy2005

    • Author(s)
      Sazaki, et al.
    • Journal Title

      Crystal Growth & Design 5

      Pages: 1729-1735

    • Related Report
      2005 Annual Research Report
  • [Journal Article] 格子欠陥発生機構の解明に向けたタンパク質結晶表面の高分解光学観察2005

    • Author(s)
      左崎 元ら
    • Journal Title

      応用物理 74

      Pages: 608-612

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Kinetic acceleration of the growth rate of glucose isomerase crystal under high pressure2005

    • Author(s)
      Suzuki, et al.
    • Journal Title

      Journal of Physical Chemistry B 109

      Pages: 3222-3226

    • Related Report
      2005 Annual Research Report
  • [Journal Article] A unique dye-decolorizing peroxidase, DyP, from Thanatephorus cucumeris Dec 1 : heterologous expression, crystallization and preliminary X-ray analysis2004

    • Author(s)
      T.Sato
    • Journal Title

      Acta Cryst. D60

      Pages: 149-152

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Novel coupling effects of the magnetic and electric fields of protein crystallization2004

    • Author(s)
      G.Sazaki
    • Journal Title

      J. Crystal Growth 262

      Pages: 499-502

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] In-situ observation of elementary growth steps on the surface of protein crystals by laser confocal microscopy2004

    • Author(s)
      G.Sazaki
    • Journal Title

      J. Crystal Growth 262

      Pages: 536-542

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] The Use of a new ad hoc growth cell with parallel electrode for the nucleation control of lysozyme2004

    • Author(s)
      A.Moreno
    • Journal Title

      J. Crystal Growth 264

      Pages: 438-444

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Effects of high pressure on the solubility and growth kinetic of monoclinic lysozyme crystals2004

    • Author(s)
      T.Asai
    • Journal Title

      Cell. Mol. Biol. 50

      Pages: 329-334

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] タンパク質結晶化への新たなアプローチ : 磁場および高圧力の利用2004

    • Author(s)
      佐崎 元
    • Journal Title

      生物物理 44

      Pages: 58-63

    • NAID

      110001158630

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] タンパク質の結晶化に及ぼす圧力効果2004

    • Author(s)
      佐崎 元
    • Journal Title

      日本結晶学会誌 46

      Pages: 152-158

    • NAID

      10012907347

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Novel coupling effects of the magnetic and electric fields on protein crystallization2004

    • Author(s)
      G.Sazaki
    • Journal Title

      J.Crystal Growth 262

      Pages: 499-502

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] In-situ observation of elementary growth steps on the surface of protein crystals by laser confocal microscopy2004

    • Author(s)
      G.Sazaki
    • Journal Title

      J.Crystal Growth 262

      Pages: 536-542

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] The Use of a new ad hoc growth cell with parallel electrodes for the nucleation control of lysozyme2004

    • Author(s)
      A.Moreno
    • Journal Title

      J.Crystal Growth 264

      Pages: 438-444

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Effects of high pressure an the solubility and growth kinetics of monoclinic lysozyme crystals2004

    • Author(s)
      T.Asai
    • Journal Title

      Cell.Mol.Biol. 50

      Pages: 329-334

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] A novel approach to protein crystallization : Use of a magnetic field and high pressure2004

    • Author(s)
      G.Sazaki
    • Journal Title

      Seibutsu Butsuri 44

      Pages: 58-63

    • NAID

      110001158630

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Effects of high pressure on protein crystallization2004

    • Author(s)
      G.Sazaki
    • Journal Title

      J.Jpn.Assoc.Cryst.Growth 46

      Pages: 152-158

    • NAID

      10012907347

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Novel coupling effects of the magnetic and electric fields on protein crystllization2004

    • Author(s)
      G.Sazaki, et al.
    • Journal Title

      J.Crystal Growth 262

      Pages: 499-502

    • Related Report
      2004 Annual Research Report
  • [Journal Article] In-situ observation of elementary growth steps on the surface of protein crstals by laser confocal microscopy2004

    • Author(s)
      G.Sazaki, et al.
    • Journal Title

      J.Crystal Growth 262

      Pages: 536-542

    • Related Report
      2004 Annual Research Report
  • [Journal Article] The Use of a new ad hoc growth cell with parallel electrodes for the nucleaion control of lysozyme2004

    • Author(s)
      A.Moreno, et al.
    • Journal Title

      J.Crystal Growth 264

      Pages: 438-444

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Effects of high pressure on the solubility and growth kinetics of monoclini lysozyme crystals2004

    • Author(s)
      T.Asai, et al.
    • Journal Title

      Cell.Mol.Biol. 50

      Pages: 329-334

    • Related Report
      2004 Annual Research Report
  • [Journal Article] タンパク質結晶化への新たなアプローチ:磁場および高圧力の利用2004

    • Author(s)
      佐崎 元ら
    • Journal Title

      生物物理 44

      Pages: 58-63

    • NAID

      110001158630

    • Related Report
      2004 Annual Research Report
  • [Journal Article] タンパク質の結晶化に及ぼす圧力効果2004

    • Author(s)
      佐崎 元ら
    • Journal Title

      日本結晶学会誌 46

      Pages: 152-158

    • NAID

      10012907347

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Step velocity in tetragonal lysozyme growth as a function of impurity concentration and mass transport conditions

    • Author(s)
      P.Dold
    • Journal Title

      J. Crystal Growth (印刷中)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Step velocity in tetragonal lysozyme growth as a function of impurity concentration and mass transport conditions

    • Author(s)
      P.Dold
    • Journal Title

      J.Crystal Growth (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] 高圧力下の生物科学 : タンパク質結晶表面のステップ前進速度に及ぼす圧力効果2006

    • Author(s)
      鈴木良尚
    • Publisher
      さんえい出版
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] 高圧力下の生物科学:タンパク質結晶表面のステップ前進速度に及ぼす圧力効果2006

    • Author(s)
      鈴木良尚ら
    • Publisher
      さんえい出版(投稿中)
    • Related Report
      2005 Annual Research Report
  • [Book] タンパク質の結晶化 : 第11章2節 磁場中で結晶をつくる2005

    • Author(s)
      佐崎 元
    • Total Pages
      11
    • Publisher
      京都大学学術出版会
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] タンパク質の結晶化 : 12章2節 タンパク質結晶の光学顕微鏡を用いた表面観察2005

    • Author(s)
      佐崎 元
    • Total Pages
      6
    • Publisher
      京都大学学術出版会
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] タンパク質の結晶化:第11章2節 磁場中で結晶をつくる2005

    • Author(s)
      佐崎 元
    • Total Pages
      11
    • Publisher
      京都大学学術出版会
    • Related Report
      2005 Annual Research Report
  • [Book] タンパク質の結晶化:12章2節 タンパク質結晶の光学顕微鏡を用いた表面観察2005

    • Author(s)
      佐崎 元
    • Total Pages
      6
    • Publisher
      京都大学学術出版会
    • Related Report
      2005 Annual Research Report
  • [Patent(Industrial Property Rights)] 光学顕微鏡と光学的観察方法2005

    • Inventor(s)
      小林 茂
    • Industrial Property Rights Holder
      小林 茂
    • Industrial Property Number
      2005-086550
    • Filing Date
      2005-03-24
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary

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Published: 2004-04-01   Modified: 2016-04-21  

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