• 제목/요약/키워드: MALDI-TOF/MS

검색결과 280건 처리시간 0.027초

Comparative proteomic analysis of plant responses to sound waves in Arabidopsis

  • Kwon, Young Sang;Jeong, Mi-Jeong;Cha, Jaeyul;Jeong, Sung Woo;Park, Soo-Chul;Shin, Sung Chul;Chung, Woo Sik;Bae, Hanhong;Bae, Dong-Won
    • Journal of Plant Biotechnology
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    • 제39권4호
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    • pp.261-272
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    • 2012
  • Environmental factors greatly influence the growth, development, and even genetic characteristics of plants. The mechanisms by which sound influences plant growth, however, remain obscure. Previously, our group reported that several genes were differentially regulated by specific frequenciesof sound treatmentusing a sound-treated subtractive library. In this study, we used a proteomic approach to investigate plant responses to sound waves in Arabidopsis. The plants were exposed to 250-Hz or 500-Hz sound waves, and total proteins were extracted from leaves 8 h and 24 h after treatment. Proteins extracted from leaves were subjected to 2-DE analysis. Thirty-eight spots were found to be differentially regulated in response to sound waves and were identified using MALDI-TOF MS and MALDI-TOF/TOF MS. The functions of the identified proteins were classified into photosynthesis, stress and defense, nitrogen metabolism, and carbohydrate metabolism. To the best of our knowledge, this is the first report on the analysis of protein changes in response to sound waves in Arabidopsis leaves. These findings provide a better understanding of the molecular basis of responses to sound waves in Arabidopsis.

Characterization of Thermal Degradation of Polytrimethylene Terephthalate by MALDI-TOF Mass Spectrometry

  • Jang, Sung-Woo;Yang, Eun-Kyung;Jin, Sung-Il;Cho, Young-Dal;Choe, Eun-Kyung;Park, Chan-Ryang
    • Bulletin of the Korean Chemical Society
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    • 제33권3호
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    • pp.833-838
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    • 2012
  • The thermal degradation products of polytrimethylene terephthalate (PTT) obtained by heating the sample in the temperature range of $250-360^{\circ}C$ under non-oxidative conditions was characterized using MALDI-TOF (matrix assisted laser desorption/ionization) mass spectrometry. The structures of the degradation products were determined and the relative compositions were estimated. The MALDI-TOF mass spectra of the thermally degraded PTT sample showed three main series of oligomer products with different end groups, which were carboxyl/carboxyl, carboxyl/allyl, and allyl/allyl. In contrast to the thermal degradation of polyethylene terephthalate (PET), the oligomers containing terephthalic anhydrides were not detected, whereas the formation of oligomers containing the unsaturated allyl ester group was confirmed by mass assignment. From these results, it was concluded that the thermal degradation of PTT proceeds exclusively through the ${\beta}$-CH hydrogen transfer mechanism, which is in accordance with the proposed reaction mechanism for the thermal degradation of polybutylene terephthalate (PBT).

Analysis of Branched PEG-Conjugated Interferon Alpha by Capillary Electrophoresis and MALDI- TOF Mass Spectrometry

  • Na, Dong-Hee;Park, Eun-Ji;Lee, Snag-Deuk;Jo, Young-Woo;Lee, Sung-Hee;Kim, Won-Bae;Lee, Kang-Choon
    • 대한약학회:학술대회논문집
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    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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    • pp.406.3-407
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    • 2002
  • Analysis of interferon alpha (IFN) modified with high molecular weight branched PEG was performed by capillary electrophoresis (CE) and MALDI-TOF mass spectrometry (MALDI-TOF MS). IFN was modified by the reaction of amine residues with an active ester of monomethoxy polyethylene glycol at various molar ratios. As a CE method. capilary electrophoresis sodium dodecyl sulfate nongel sieving (CE-SDS NGS) was performed using an uncoated capilary filled with a hydrophilic replaceable polymer network matrix. (omitted)

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고효율의 ESI-TOF/Ms 시스템을 이용한 생리활성 항체와 방사성동위원소 표지용 착화제의 결합 검증 (Verification with of High Efficiency Chemical Binding System of a Physiologically Active Radioisotope Using ESI-TOF/Ms System)

  • 조은하;홍영돈;최선주
    • 약학회지
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    • 제57권6호
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    • pp.400-405
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    • 2013
  • In this study, we measured the complex efficiency of a physiologically active antibody, a chelator and radiosiotopes using the ESI-TOF/Ms system for develop good radiopharmaceuticals. For a precise measurement, TLC is a low accuracy method. Loading of same amount of sample is difficult for each test, and work to quantify accurately the results obtained through TLC cannot be afforded compared to the use of other analytical instruments. The method of analysis using a mass spectrometer is capable of a mass analysis of proteins for quantitative analysis. The conjugates of the chelator (CHX-A- DTPA) and the antibody (IgG) were separated through MWCO, and were analyzed using ESI-TOF and MALDI-TOF mass spectrometry. The analysis using MALDI-TOF is roughly divided into measurements on mass spectrometry. When conjugating a small molecular weight of CHX-A-DTPA and a large molecular weight of IgG, distinguishing the peak of the conjugate and the peak of IgG was difficult. However, an ESI-TOF mass spectrometer system is capable of an analysis of mass by decentralizing the IgG. It is utilized as a technique for measuring the metabolic processes during conjugation and the stability evaluation of radiopharmaceuticals. When establishing this technique, the accuracy of the overall radiophar-maceutical analysis is expected to be able to be improved.

Application of Malononitrile Derivatization Method for Structural Glycomics Study in Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry

  • Ahn, Yeong-Hee;Yoo, Jong-Shin
    • Journal of Photoscience
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    • 제8권2호
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    • pp.83-86
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    • 2001
  • Structural analyses of oligosaccharide-malononitrile derivatives were conducted by matrix-assisted laser desorption/ionization post-source decay (MALDI-PSD) analysis in positive ion mode. The malononitrile derivatives of oligosaccharides, which were developed for highly sensitive detection of multi-component oligosaccharides by negative ion electrospray ionization mass spectrometry (ESI MS), were detected by positive-ion MALDI with the detection limit of 2 pmol level from the crude derivatization sample. The used matrix affected drastically the analytical results of oligosaccharide-malononitrile derivative by matrix-assisted laser desoprtion/ionization mass spectrometry (MALDI MS). The malononitrile derivatization of oligosaccharide also affect the patterns of MALDI-PSD spectra and give much more structural information than the free oligosaccharide.

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Proteome analysis of chloroplast proteins in stage albinism line of winter wheat (triticum aestivum) FA85

  • Hou, Dian-Yun;Xu, Hong;Du, Guang-Yuan;Lin, Jun-Tang;Duan, Min;Guo, Ai-Guang
    • BMB Reports
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    • 제42권7호
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    • pp.450-455
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    • 2009
  • The "stage albinism line of winter wheat" FA85 was a specific natural mutant strain on leaf color. This physiological mutation was controlled by cytogene. In order to reveal the genetic and biochemical mechanism of albinism, 2-DE was used to investigate the difference of chloroplast protein expression pattern between FA85 and its parent wheat Aibian 1. From the results of 2-DE gels analysis, approximately 683 spots were detected on each gel, and 57 spots were expressed differently at least two-fold. Using MALDI-TOF/TOF MS, 14 of 57 spots were identified, which could be categorized into four classes: carbon metabolism, energy metabolism, defense/stress response and signal transduction. Compared with the parent wheat, the expression of ATPase-$\gamma$ and GP1-$\alpha$ was up-regulated in FA85, and of other proteins was down-regulated. Together, we concluded that the expression of chloroplast proteins had changed obviously in FA85, which might be related to the leaf color mutant.

추백리가 감염된 닭의 간에서 발현이 증가하는 APOA1 단백질의 확인 (Identification of Upregulated APOA1 Protein of Chicken Liver in Pullorum Disease)

  • 정기철;이유주;유성란;이준헌;장병귀;구용범;소현경;최강덕
    • 한국가금학회지
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    • 제32권1호
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    • pp.23-27
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    • 2005
  • 가금 추백리와 밀접한 관계가 있는 단백질을 동정하기 위하여 인위적으로 추백리를 유도한 개체와 정상인 개체의 간에서 단백질을 추출하였으며, 2차원 전기영동(2DE)과 mass spectrometry(MS)를 이용하여 차등 발현되는 단백질을 조사하였다. 총 300여개의 단백질 spot들이 관찰되었으며, 이중 발현양에 현저한 차이를 보이는 spot을 MALDI-TOF MS와 protein database search를 통해 분석한 결과, 가금 APOAI (Apolipoprotein A1)으로 확인되었다. 추백리가 감염된 닭의 간에서 APOA1 단백질의 증가는 손상된 혈관의 재생과 밀접한 관계가 있으며 추백리의 감염 여부를 나타내 주는 Bio-marker로서 활용될 수 있을 것으로 사료된다.

인삼 모상근 프로테옴 데이터 분석 : 인삼 EST database와의 통합 분석에 의한 단백질 동정 (Proteome Data Analysis of Hairy Root of Panax ginseng : Use of Expressed Sequence Tag Data of Ginseng for the Protein Identification)

  • 권경훈;김승일;김경욱;김은아;조건;김진영;김영환;양덕춘;허철구;유종신;박영목
    • Journal of Plant Biotechnology
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    • 제29권3호
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    • pp.161-170
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    • 2002
  • 인삼 모상근의 프로테옴 분석에 의해 얻은 질량분석 스펙트럼 데이터는 MALDI/TOF/MS에서 얻는 질량 스펙트럼과 ESI/Q-TOF/MS에서 얻는 탄뎀 질량 스펙트럼으로 구분된다. 질량 스펙트럼은 단백질이 효소에 의해 분해된 펩타이드들의 분자량 정보를 제공하며, 탄뎀 질량 스펙트럼에서는 아미노산 단위로 분해된 절편 단백질의 분자량으로부터 아미노산 서열을 결과로 얻는다. 펩타이드의 아미노산 서열을 BLAST로 검색하면 유사한 단백질을 GenBank에서 검색할 수 있다. 이러한 단백질 동정 방법은 완전한 유전체 서열이 알려진 생물체의 경우 높은 정확도로 단백질을 동정할 수 있으나, 그렇지 않은 경우는 유사한 단백질이 데이터베이스에 존재하지 않아 분석이 용이하지 않다. 본 연구에서는 질량 스펙트럼 및 절편 단백질의 아미노산 서열을 EST (expressed sequence tag) 서열과 비교하여 프로테옴 데이터와 일치하는 EST 서열을 찾아내고 이를 BLAST검색에 의해 단백질 동정에 활용하였다. ESI/Q-TOF/MS 에서 얻은 아미노산 서열은 길이는 짧지만 데이터의 신뢰도가 높으므로 EST 서열과의 연관 관계를 밝힘으로써 단백질에 대한 정보를 보완할 수 있었다. ESI/Q-TOF/MS에서 얻은 펩타이드의 아미노산 서열을 EST 서열과 비교한 결과 90%의 아미노산 서열이 EST DB에서 발견되었다. NCBI의 nr 데이터베이스에서 아미노산 서열을 검색하여 찾은 단백질이 68%임에 비하여, 인삼 EST 서열에 의한 검색이 22% 더 많은 결과를 얻었다. MALDI/TOF/MS의 질량 스펙트럼에서 nr 데이터베이스로 검색한 결과와 인삼 EST 데이터베이스를 검색한 결과가 일치하는 경우는 47개 중 9개인 19%에 불과하여, 탄뎀 질량 분석으로 아미노산 서열을 얻지 않고, 단지 질량 스펙트럼으로부터 단백질을 동정하는 방법으로는 단백질 동정의 정확한 결과를 기대하기 어려움을 확인하였다.

IDENTIFICATION OF DIFFERENTIALLY EXPRESSED PROTEINS IN DIFFERENT GROWING STAGES IN CHICKEN LIVER BY PROTEOMICS APPROACH

  • Lee, K.Y.;Jung, K.C.;Jang, B.G.;Choi, K.D.;Lee, J.H.
    • 한국가금학회:학술대회논문집
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    • 한국가금학회 2006년도 제23차 정기총회 및 학술발표회
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    • pp.74-76
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    • 2006
  • 닭의 간은 해독작용, 당의 저장, 혈장 단백질의 합성 등 주요 기능을 하는 것으로 알려져 있다. 본 연구는 간에서 성장 단계별로 발현량에 차이를 보이는 단백질들을 비교해 보았다. 2차원적 전기 영동에 의해 분리된 300개 이상의 단백질들이 확인되었으며 이 중 성장 단계별 특이적인 13개의 단백질은 MALDI-TOF MS에 의하여 분석이 되어졌다. 본 연구를 통하여 밝혀진 단백질들은 생화학적인 연구에 중요한 자료를 제공할 것으로 사료된다.

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