• 제목/요약/키워드: matrix assisted laser desorption/ ionization (MALDI)

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Matrix-assisted Laser Desorption/ Ionization Time-of-flight Mass Spectrometry를 이용한 화장품에서의 계면활성제 분석

  • 이명희;김상진
    • 대한화장품학회지
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    • 제25권3호
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    • pp.1-21
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    • 1999
  • 화장품, 의료용, 가정용품 및 공업용제품에까지 광범위한 용도로 사용량이 많은 중요한 비이온 계면활성제의 종류 중에서 polyoxyethylene(POE) 부가형 계면활성제의 경우 그 부가몰수에 따라 특성이 달라지고 용도가 다르게 사용된다. 이때 부가된 분자들의 분자량이 크고 분포를 이루는 혼합물이기 때문에 분석이 매우 어렵다. 따라서 MALDI-TOF/MS 방법을 이용하여 이들의 분자량과 그 분포를 측정함으로써 쉽고 빠르게 측정할 수 있는 새로운 방법을 개발하고자 하였다. 이 논문에서는 화장품에 주로 사용되는 비이온 계면활성제를 선택하여 MALDI-TOF/AfS를 측정하여 스펙트럼으로부터 분자량 분포와 POE부가정도를 측정할 수 있었다. 그리고 이 조건을 적용하여 시중에 판매되는 제품에서 추출된 비이온 계면활성제의 MALDI-TOF/MS 스펙트럼으로부터 분자량 분포와 POE 부가 정도를 확인 할 수 있었다.

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Quantitative Proteomics Towards Understanding Life and Environment

  • Choi, Jong-Soon;Chung, Keun-Yook;Woo, Sun-Hee
    • 한국환경농학회지
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    • 제25권4호
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    • pp.371-381
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    • 2006
  • New proteomic techniques have been pioneered extensively in recent years, enabling the high-throughput and systematic analyses of cellular proteins in combination with bioinformatic tools. Furthermore, the development of such novel proteomic techniques facilitates the elucidation of the functions of proteins under stress or disease conditions, resulting in the discovery of biomarkers for responses to environmental stimuli. The ultimate objective of proteomics is targeted toward the entire proteome of life, subcellular localization biochemical activities, and the regulation thereof. Comprehensive analysis strategies of proteomics can be classified into three categories: (i) protein separation via 2-dimensional gel electrophoresis (2-DE) or liquid chromatography (LC), (ii) protein identification via either Edman sequencing or mass spectrometry (MS), and (iii) proteome quantitation. Currently, MS-based proteomics techniques have shifted from qualitative proteome analysis via 2-DE or 2D-LC coupled with off-line matrix assisted laser desorption ionization (MALDI) and on-line electrospray ionization (ESI) MS, respectively, toward quantitative proteome analysis. In vitro quantitative proteomic techniques include differential gel electrophoresis with fluorescence dyes. protein-labeling tagging with isotope-coded affinity tags, and peptide-labeling tagging with isobaric tags for relative and absolute quantitation. In addition, stable isotope-labeled amino acids can be in vivo labeled into live culture cells via metabolic incorporation. MS-based proteomics techniques extend to the detection of the phosphopeptide mapping of biologically crucial proteins, which ale associated with post-translational modification. These complementary proteomic techniques contribute to our current understanding of the manner in which life responds to differing environment.

Proteome Analysis of Waito-c Rice Seedlings Treated with Culture Fluid of Gibberellin-producing Fungus, Fusarium proliferatum KGL0401

  • Rim, Soon-Ok;Lee, Jin-Hyung;Hwang, Seon-Kap;Suh, Seok-Jong;Lee, Jin-Man;Rhee, In-Koo;Kim, Jong-Guk
    • Journal of Microbiology and Biotechnology
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    • 제16권12호
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    • pp.1990-1994
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    • 2006
  • Fusarium proliferatum KGL0401 was previously isolated from Physalis alkekengi var. francheti plant roots and exhibited a high GA productivity. A gas chromatography-mass spectrometry (GC-MS) analysis of extracts of the culture fluid of F proliferatum KGL0401 also revealed the presence of $GA_1$, $GA_3$, $GA_4$, $GA_7$, $GA_{20}$, and $GA_{24}$. Therefore, the present study conducted a proteome analysis of waito-c rice treated with the culture fluid of the isolated F proliferatum KGL0401 to identify the protein expression triggered by the GA-containing culture fluid. The results revealed the overexpression of 180 protein spots in the sample treated with the culture fluid. Among them, 75 induced proteins were selected and analyzed by MALDI-TOF (matrix-assisted laser desorption-iorrization time-of-flight) mass spectrometry, followed by database searching, and 51 proteins were identified.

Investigation of Angiotensin Glycosylation by MALDI-TOF and ESI Tandem Mass Spectrometry

  • Park, Soo-Jin;Park, Deok-Hie;Sul, Soo-Hwan;Oh, Sung-Hwan F.;Park, In-Sook;Chung, Doo-Soo;Kim, Hie-Joon;Kim, Min-Sik;Lee, Sang-Won
    • Bulletin of the Korean Chemical Society
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    • 제25권12호
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    • pp.1791-1800
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    • 2004
  • Angiotensin I, a model decapeptide, was glycosylated and partially hydrolyzed with HCl (6 N, 80 $^{\circ}C$, 4 h), aminopeptidase, and carboxypeptidase Y. A single peptide mass map obtained from truncated peptides in the partial acid hydrolysate of angiotensin and its glycosylation product mixture by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry enabled sequencing of angiotensin by a combinatorial procedure. MALDI-TOF and electrospray ionization (ESI) tandem mass spectrometric results indicate that both the N-terminal amino group of aspartic acid and the guanidinium group of the second residue arginine are glycosylated.

Application of MALDI Tissue Imaging of Drugs and Metabolites: A New Frontier for Molecular Histology

  • Shanta, Selina Rahman;Kim, Young-Jun;Kim, Young-Hwan;Kim, Kwang-Pyo
    • Biomolecules & Therapeutics
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    • 제19권2호
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    • pp.149-154
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    • 2011
  • Matrix assisted laser desorption ionization (MALDI) mass spectrometry is commonly used to analyze biological molecules such as proteins, peptides and lipids from cells or tissue. Recently MALDI Imaging mass spectrometry (IMS) has been widely applied for the identification of different drugs and their metabolites in tissue. This special feature has made MALDI-MS a common choice for investigation of the molecular histology of pathological samples as well as an important alternative to other conventional imaging methods. The basic advantages of MALDI-IMS are its simple technique, rapid acquisition, increased sensitivity and most prominently, its capacity for direct tissue analysis without prior sample preparation. Moreover, with ms/ms analysis, it is possible to acquire structural information of known or unknown analytes directly from tissue sections. In recent years, MALDI-IMS has made enormous advances in the pathological field. Indeed, it is now possible to identify various changes in biological components due to disease states directly on tissue as well as to analyze the effect of treated drugs. In this review, we focus on the advantages of MALDI tissue imaging over traditional methods and highlight some motivating findings that are significant in pathological studies.

임상검체에서 분리된 Ochrobactrum anthropi의 효과적인 동정 (Effective Identification of Ochrobactrum anthropi Isolated from Clinical Specimens)

  • 고현미;조준현;백해경
    • 대한임상검사과학회지
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    • 제52권3호
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    • pp.221-228
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    • 2020
  • Ochrobactrum anthropi는 oxidase를 생산하는 비발효산소성 그람음성 막대균으로 외관이 비슷하고 oxidase가 양성인 비발효 세균과 혼합배양 시 구분이 힘들고 생화학적 동정 장비로는 정확한 동정에 한계가 있다. 따라서 본 연구에서는 생화학적 검사 방법으로 동정이 힘든 세균동정의 Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Platform (MALDI-TOF) 법의 유용성을 제시하고자 하였다. MicroScan을 이용해 검사했던 O. anthropi 5례를 분석한 결과, 최종보고까지 6.2일이 소요되었으며 E. coli의 3.0일에 비해 3.5일이 더 소요되었다. 5번 환자 고름 검체는 Achromobater xylosoxidans와 혼합감염으로 여러 번의 계대 배양과 재검사로 인해 11.3일이 소요되었는데, MALDI-TOF법으로 검사한 경우 한 번에 동정되었다. 4명의 환자는 기저질환이 있는 60세 이상이었고 기회감염과 원내감염의 가능성을 배제할 수 없었으며, 그 중 92일 만에 채취된 검체는 imipenem과 meropenem에 내성이었다. 따라서 O. anthropi처럼 동정이 까다로운 세균은 신속하고 적절한 환자 치료를 위해 MALDI-TOF법을 이용한 검사가 매우 유용할 것으로 사료된다.

A MALDI-MS-based Glucan Hydrolase Assay Method for Whole-cell Biocatalysis

  • Ahn, Da-Hee;Park, Han-Gyu;Song, Won-Suk;Kim, Seong-Min;Jo, Sung-Hyun;Yang, Yung-Hun;Kim, Yun-Gon
    • 한국미생물·생명공학회지
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    • 제47권1호
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    • pp.69-77
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    • 2019
  • Screening microorganisms that can produce glucan hydrolases for industrial, environmental, and biomedical applications is important. Herein, we describe a novel approach to perform glucan hydrolase screening-based on analysis of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) spectra-which involves degradation of the oligo- and polysaccharides. As a proof-of-concept study, glucan hydrolases that could break down glucans made of several glucose units were used to demonstrate the MALDI-MS-based enzyme assay. First, the enzyme activities of ${\alpha}$-amylase and cellulase on a mixture of glucan oligosaccharides were successfully discriminated, where changes of the MALDI-MS profiles directly reflected the glucan hydrolase activities. Next, we validated that this MALDI-MS-based enzyme assay could be applied to glucan polysaccharides (i.e., pullulan, lichenan, and schizophyllan). Finally, the bacterial glucan hydrolase activities were screened on 96-well plate-based platforms, using cell lysates or samples of secreted enzyme. Our results demonstrated that the MALDI-MS-based enzyme assay system would be useful for investigating bacterial glucoside hydrolases in a high-throughput manner.

Comparative Proteome Analysis of Celastrol-Treated Helicobacter pylori

  • Kim, Sa-Hyun
    • 대한의생명과학회지
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    • 제23권4호
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    • pp.395-401
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    • 2017
  • Various preclinical and clinical trials have been conducted the efficacy of celastrol. In data presented in the current manuscript is the first trial to inhibit Helicobacter pylori with celastrol. In this study, the quantitative change of various H. pylori proteins including CagA and VacA by the anti-bacterial effect of celastrol was determined. The anti-H. pylori effects of celastrol was investigated by performing 2-dimensional electrophoresis and additional supporting experiments. After 2-dimensional electrophoresis analysis, spot intensities were analyzed and then each spot was identified using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) or peptide sequencing using Finnigan LCQ ion trap mass spectrometer (LC-MS/MS). The results show that celastrol has multiple effects on protein expression in H. pylori.

Postharvest Biological Control of Colletotrichum acutatum on Apple by Bacillus subtilis HM1 and the Structural Identification of Antagonists

  • Kim, Hae-Min;Lee, Kui-Jae;Chae, Jong-Chan
    • Journal of Microbiology and Biotechnology
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    • 제25권11호
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    • pp.1954-1959
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    • 2015
  • Bacillus subtilis HM1 was isolated from the rhizosphere region of halophytes for its antifungal activity against Colletotrichum acutatum, the causative agent of anthracnose. Treatment of postharvest apples with the cell culture or with a cell-free culture supernatant reduced disease severity 80.7% and 69.4%, respectively. Both treatments also exhibited antifungal activity against various phytopathogenic fungi in vitro. The antifungal substances were purified and analyzed by acid precipitation, gel filtration, high-performance liquid chromatography, and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Three compounds were identified as fengycin, iturin, and surfactin. The MALDI-TOF/TOF mass spectrum revealed the presence of cyclized fengycin homologs A and B, which were distinguishable on the basis of the presence of either alanine or valine, respectively, at position 6 of the peptide sequence. In addition, the cyclized structure of fengycin was shown to play a critical role in antifungal activity.

Protein Expression Profiling of Infected Murine Macrophage Cells (RAW 264.7) by Bacillus anthracis Spores

  • Seo Gwi-Moon;Nam Jeong-Ah;Oh Kwang-Gun;Chai Young-Gyu
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 2003년도 International Meeting of the Microbiological Society of Korea
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    • pp.77-79
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    • 2003
  • Current therapeutic strategies far anthrax have had no significant impact on anthrax mortality over the last several decades. This study used a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) discovery platform to generate protein expression profiles in search of overexpressed proteins in murine macrophage cells (RAW264.7) which infected with Bacillus anthracis spores as potentially novel molecular targets. Two differentially expressed proteins were identified in infected murine macrophage cells as Syndapin and CDC46, respectively. Syndapins are potential links between the cortical actin cytoskeleton and endocytosis. Other two proteins were identified from murine macrophage cells infected with avirulent spores as ITBG-2 (CD18) and HSPA5, respectively. These data demonstrate the feasibility of using a MALDI-TOF platform to generate protein expression profiles and identify potential molecular targets for anthrax therapeutics.

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