• 제목/요약/키워드: Protein Crystallization

검색결과 52건 처리시간 0.023초

Immunological Analysis of Antigenic Variation of Bacillus thuringiensis subsp. sotto during Sporulation and Crystallization

  • Cho, Jae Min;Gi Bum Nam;Soon Bok Hong;Myung Hwan Cho
    • Journal of Microbiology and Biotechnology
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    • 제5권6호
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    • pp.359-363
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    • 1995
  • The antigenic variation of B. thuringiensis subsp. satto have been investigated for 120 hours during sporulation and crystallization by using SDS-PAGE and Western blot. Most antigens of a vegetative cell were found to disappear as it was in sporulation and crystallization, but protein antigens of 46, 29, 27, and 21 kDa continued to be expressed. The new protein bands of 293, 138, 119, 75, and 68 kDa appeared on days 2 through 5 in modified GYS medium. They were thought to be involved in sporulation and crystallization. The protein of 138 kDa was found to be a major protein of both crystal and spore. The expression patterns were immunologically analyzed by Western blot. The polyclonal antisera against the intact crystal showed strong immunoreactivity to proteins with molecular masses of 293, 138, 68, and 46 kDa. The polyclonal antisera against the spore recognized proteins of 293, 138, 68, and 46 kDa. Both crystals and spores appeared to express the common protein antigens.

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Development of Crystallization Distinction Supporting System Using Image Processing

  • Saito, Kanako;Kawabata, Kuniaki;kunimitsu, Satoshi;Asama, Hajime;Mishima, Taketoshi
    • 대한전자공학회:학술대회논문집
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    • 대한전자공학회 2002년도 ITC-CSCC -3
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    • pp.1788-1791
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    • 2002
  • In the post-genome era. it is one of important research subject to Investigate the roles of the proteins in human body based on decoded genome information during Human Genome Project. In order to clarify them. it is necessary to analyze the structure of the protein crystals and their function. ' Crystallization is the beginning stage of protein structure determination process. There are some methods for structural analysis of the proteins, and general one is X-ray structural analysis method. In order to utilize this method for analyzing the protein crystal's structure, artificial protein crystallization is required. However, since artificial crystallizing work takes much time and manpower. the performance against its cost is still low. Therefore. we started to discuss to develop a supporting system for improving efficiency of the crystallization distinction procedure. In this paper, we examine to realize such supporting system for crystallization distinction using image-processing technique and report about our experimental result with many real protein solution images.

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Crystallization of amorphous Si by pulse annealing with Ni ferritins

  • Tojo, Yosuke;Miura, Atsushi;Fuyuki, Takashi;Yamashita, Ichiro;Uraoka, Yukiharu
    • 한국정보디스플레이학회:학술대회논문집
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    • 한국정보디스플레이학회 2009년도 9th International Meeting on Information Display
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    • pp.553-556
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    • 2009
  • We investigated an application of supramolecular protein, and demonstrated the metal induced lateral crystallization utilizing ferritins with Ni nanoparticles, named the "bio-nano-crystallization". So far, this method has required long time, because of this method condition based on the conventional solid phase crystallization. In this study, we applied the pulsed rapid thermal annealing to bio-nanocrystallization. As a result, we succeeded in the crystallization for a short time. We found that the TFTs characteristics were improved with decrease metal impieties in poly-Si thin films by this method.

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Application of hybrid LRR technique to protein crystallization

  • Jin, Mi-Sun;Lee, Jie-Oh
    • BMB Reports
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    • 제41권5호
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    • pp.353-357
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    • 2008
  • LRR family proteins play important roles in a variety of physiological processes. To facilitate their production and crystallization, we have invented a novel method termed "Hybrid LRR Technique". Using this technique, the first crystal structures of three TLR family proteins could be determined. In this review, design principles and application of the technique to protein crystallization will be summarized. For crystallization of TLRs, hagfish VLR receptors were chosen as the fusion partners and the TLR and the VLR fragments were fused at the conserved LxxLxLxxN motif to minimize local structural incompatibility. TLR-VLR hybridization did not disturb structures and functions of the target TLR proteins. The Hybrid LRR Technique is a general technique that can be applied to structural studies of other LRR proteins. It may also have broader application in biochemical and medical application of LRR proteins by modifying them without compromising their structural integrity.

Evaporation Rate in Protein Crystallization Via Vapor Diffusion can be Controlled through a Simple Multistep-concentration Setting in Capillaries

  • Lee, Min-Nyung;Chung, Yong-Je
    • 한국결정학회지
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    • 제14권1호
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    • pp.35-38
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    • 2003
  • 증기-확산 단백질 결정화의 물-증발 속도를 조절하기 위해서, 간결한 모세관 내의 다중-단계 농도 장착법 (multistep-concentration setting in capillaries) 이 이용되었다. “조절 용액 (regulatory solution)”이라고 일컬어지는 2차 침전 용액이 이용되어, 이 방법으로 다양한 증발 속도 곡선들이 얻어졌다. 이때, 조절 용액은 단백질 용액에 직접적으로 노출되지 않는다. 모델 단백질인 lysozyme의 결정화에 이 그래프들이 적용되었다. 결정 성장은 증발속도에 달려있다는 것을 실험 결과들이 명백하게 보여주었다. 특히나. 단백질 용액의 침전 농도가 어떤 점까지 증가하다가 평형 농도로 줄어드는 decoupling 곡선이 가장 좋은 결정들을 만들어냈다.

Coexpression and protein-protein complexing of DIX domains of human Dvl1 and Axin1 protein

  • Choi, Seung-Hye;Choi, Kyung-Mi;Ahn, Hyung-Jun
    • BMB Reports
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    • 제43권9호
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    • pp.609-613
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    • 2010
  • The Dvl and Axin proteins, which are involved in the Wnt signaling pathway, each contain a conserved DIX domain in their sequences. The DIX domain mediates interaction between Dvl and Axin, which together play an important role in signal transduction. However, the extremely low production of DIX domain fragments in E. coli has prevented more widespread functional and structural studies. In this study, we demonstrate that the DIX domains of Dvl and Axin are expressed noticeably in a multi-cistronic system but not in a mono-cistronic system. Formation of the $DIX_{Dvl1}-DIX_{Axin1}$ complex was investigated by affinity chromatography, SEC and crystallization studies. Unstable DIX domains were stabilized by complexing with counterpart DIX domains. The results of the preliminary crystallization and diffraction of the $DIX_{Dvl1}-DIX_{Axin1}$ complex may prove useful for further crystallographic studies.

재조합 GG1234-DsRed 융합 단백질의 생산 및 In vitro 탄산칼슘 결정화에 미치는 영향에 대한 연구 (Production of Recombinant GG1234-DsRed Fusion Protein and Its Effect on in vitro CaCO3 Crystallization)

  • 손채연;김진호;김지하;최유성
    • KSBB Journal
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    • 제30권6호
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    • pp.296-301
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    • 2015
  • Eggshell-based biocomposites have become attractive due to their exquisite nanostructure and biological properties, which are mainly composed of highly organized calcium carbonate crystals controlled by organic macromolecules such as proteins and polysaccharides. Here, we designed the recombinant fusion protein of a putative eggshell matrix protein named as GG1234 and a fluorescent reporter protein of DsRed. The protein was successfully over-expressed in E. coli and purified by Ni-NTA affinity chromatography. In vitro calcium carbonate crystallization was conducted in the presence of the fusion protein, and morphological change was investigated. The protein inhibited the calcite growth in vitro, and spherical calcium carbonate micro-particles with the diameter of about $20-30{\mu}m$ were obtained. We expect that this study would be helpful for better understanding of eggshell-based biomineralization.

Fe65단백질의 한 PTB 도메인에 대한 과발현 및 초기 결정화 (High-level production and initial crystallization of a Fe65 PTB domain)

  • 노승현;하남출
    • 생명과학회지
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    • 제17권1호
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    • pp.18-23
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    • 2007
  • 신경세포에 특이적으로 발현되는 단백질인 Fe65는 두 개의 phosphotyrosine binding(PTB) 도메인을 가지고 있다. 두번째 PTB(PTB2)도메인은 아밀로이드 베타 전구 단백질(APP)의 세포질 도메인 조각(AICD)와 결합한다. 최근 연구 결과들은 AICD와 Fe65로 이루어진 결합체가 알츠하이머병에서 신경세포를 죽게 하는 유전자를 발현한다고 제시하고 있다. 따라서 Fe65와 AICD의 결합을 방해하는 화합물은 알츠하이머병을 치료하는데 후보물질이 될 수 있다. 하지만 AICD와 Fe65와 관련된 신호전달에 대한 분자적 기전은 잘 알려져 있지 않다. 이번 연구에서는 Fe65의 PTB2도메인을 baculovirus시스템에서 과발현시킨 결과를 보고한다. 세균 및 척추동물 세포를 이용한 시스템과 비교했을 때, baculovirus 시스템 이 훨씬 효과적 이 다는 것을 발견했다. 정제된 재조합 단백질을 이용하여 초기 결정을 얻었다. 결정을 이용하여 앞으로 밝힐 3차원 구조는 Fe65관련 신호전달체계에 대한 분자 기전 및 이에 대한 저해제 개발에 큰 도움을 줄 것이다.

Bioinformatic approaches for the structure and function of membrane proteins

  • Nam, Hyun-Jun;Jeon, Jou-Hyun;Kim, Sang-Uk
    • BMB Reports
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    • 제42권11호
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    • pp.697-704
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    • 2009
  • Membrane proteins play important roles in the biology of the cell, including intercellular communication and molecular transport. Their well-established importance notwithstanding, the high-resolution structures of membrane proteins remain elusive due to difficulties in protein expression, purification and crystallization. Thus, accurate prediction of membrane protein topology can increase the understanding of membrane protein function. Here, we provide a brief review of the diverse computational methods for predicting membrane protein structure and function, including recent progress and essential bioinformatics tools. Our hope is that this review will be instructive to users studying membrane protein biology in their choice of appropriate bioinformatics methods.

PCL/PCL-g-PEG 생분해성 블렌드에서 그래프트 공중합체의 조성에 따른 상용성의 영향 (Effect of Graft Copolymer Composition on the Compatibility of Biodegradable PCL/PCL-g-PEG Blend)

  • 조국영;이기석;박정기
    • 폴리머
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    • 제33권3호
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    • pp.248-253
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    • 2009
  • 의료용으로 적용될 수 있는 새로운 재료를 제조하기 위하여 폴리카프로락톤(PCL)과 양친성 구조를 갖는 폴리(에틸렌 글리콜)(PEG)이 그래프트된 PCL을 이용하여 생분해성 블렌드를 제조하였다. 제조된 블렌드는 PCL을 기본으로 하고 여기에 그래프트 공중합체의 함량을 변화시키며 열적 그리고 결정화 특성을 관찰하였다. 그래프트 공중합체의 함량 변화에 따라 결정화 온도의 변화 및 결정화 속도가 변화하였고 이를 통해 그래프트 공중합체가 PCL의 결정화 거동에 영향을 미침을 확인하였다. 이는 광학현미경을 통한 결정의 교대 소광 밴드의 관찰을 통하여서도 확인할 수 있었다. 약물방출시스템과 같은 의료용 응용을 고려하여 블렌드 필름의 흡수거동과 단백질 흡착에 대한 특성도 평가하였다.